http://2007.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=250&target=CelineYZ&year=&month=2007.igem.org - User contributions [en]2024-03-29T15:22:39ZFrom 2007.igem.orgMediaWiki 1.16.5http://2007.igem.org/wiki/index.php/Celine_ZengCeline Zeng2008-04-08T03:47:58Z<p>CelineYZ: </p>
<hr />
<div>Celine is in her third year of Specialization Pharmacology. She is interested in technology commercialization and translational research; global health; international standards on traditional herbal Chinese medicine. Celine enjoys traveling, yoga, dancing, reading and swimming. She has also worked as a summer student at the Cross Cancer Institute. She is also proud to be a ButaNerd. <br><br />
<br />
Back to [[Alberta]]<br><br />
Back to [[Alberta/Members]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Celine_ZengCeline Zeng2007-12-29T02:28:45Z<p>CelineYZ: </p>
<hr />
<div>Celine is in her third year of Honors Pharmacology. She hopes to get into an Industrial Internship Program in the pharmaceutical industry after third year. After graduation, she will eventually pursue a PhD in Biomedical Sciences or Chemical Biology. Celine is interested in technology commercialization and translational research. She is also interested in influencing policy on the quality and efficacy control of traditional herbal Chinese medicine on a global level. Her ultimate career goal is to establish her own consulting and research firm ''Zenomics''. <br />
<br />
Outside of the lab, Celine enjoys traveling, yoga, dancing, reading and swimming. She is also working as a summer student at the Cross Cancer Institute. She is also proud to be a ButaNerd. <br><br />
<br />
Back to [[Alberta]]<br><br />
Back to [[Alberta/Members]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-27T04:48:37Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichiometric air to fuel ratio at 4 different angular speeds. The experimental setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomass into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum'' (i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli'''s tolerance to solvents such as butanol.<br />
<br />
I: Transforming genes in ''C. acetobutylicum'''s butanoate pathway into ''E.coli''<br><br />
Genes encoding the enzymes in the ''C. acetobutylicum'' butanoate metabolism pathway were identified using the KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy is to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
After receiving the commercially synthesized coding sequences of the genes of interest, the coding sequences are restricted with proper BioBrick prefixes and suffixes out of the original plasmid and cloned into B0034 (ribosome binding site). Double digest (Xba I and Spe I) as well as automated sequencing reactions are performed to verify the proper insertion of the coding sequences and the presence of a ribosome binding site at the 5' end of each coding sequence. <br><br />
<br />
The operon, shown below, consists of all of the genes in the pathway (except ''E. coli'''s thiolase). Note that we purposedly added a Histamine tag to the carboxy terminus of each gene product such that we could use Nickel-NTA column to purify the tagged proteins and subsequently analyze protein expression and enzymatic activity.<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
II: Developing butanol tolerance with the mutagenic and toxic compound--Ethylnitrosourea ENU<br><br />
The concept is to make butanol-agar plates of various butanol concentrations and place an ENU disk in the centre. We expect two ring-shaped kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the outer edge of the butanol-agr plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cells that would be mutated in a benifical way to render them resistant to butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the possibiilty of introducing the butanol production pathway into a photoautotrophic bacterium, ''Chlorobium tepidum''. ''C. tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''C. tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. In addition, the genome of this bacterium has been completely sequenced. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered three factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''. <br><br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - Butyryl CoA dehydrogenase<br><br />
Enny - Enoyl CoA hydratase<br><br />
Buddy - Butanol dehydrogenase<br><br />
Betty - Beta-hydraoxy butyryl CoA dehydrogenase<br><br />
Deisel Blaze - Butyraldehyde dehydrogenase<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
Meetings (Agenda's, Minutes, Action Items):<br />
<br />
[[Alberta/Calender/Meetings|Meeting Information]]<br />
<br />
<br />
Online Form:<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-27T04:45:48Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichiometric air to fuel ratio at 4 different angular speeds. The experimental setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomass into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum'' (i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli'''s tolerance to solvents such as butanol.<br />
<br />
I: Transforming genes in ''C. acetobutylicum'''s butanoate pathway into ''E.coli''<br><br />
Genes encoding the enzymes in the ''C. acetobutylicum'' butanoate metabolism pathway were identified using the KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy is to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
After receiving the commercially synthesized coding sequences of the genes of interest, the coding sequences are restricted with proper BioBrick prefixes and suffixes out of the original plasmid and cloned into B0034 (ribosome binding site). Double digest (Xba I and Spe I) as well as automated sequencing reactions are performed to verify the proper insertion of the coding sequences and the presence of a ribosome binding site at the 5' end of each coding sequence. <br><br />
<br />
The operon, shown below, consists of all of the genes in the pathway (except ''E. coli'''s thiolase). Note that we purposedly added a Histamine tag to the carboxy terminus of each gene product such that we could use Nickel-NTA column to purify the tagged proteins and subsequently analyze protein expression and enzymatic activity.<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
II: Developing butanol tolerance with the mutagenic and toxic compound--Ethylnitrosourea ENU<br><br />
The concept is to make butanol-agar plates of various butanol concentrations and place an ENU disk in the centre. We expect two ring-shaped kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the outer edge of the butanol-agr plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cells that would be mutated in a benifical way to render them resistant to butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered three factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''. <br><br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - Butyryl CoA dehydrogenase<br><br />
Enny - Enoyl CoA hydratase<br><br />
Buddy - Butanol dehydrogenase<br><br />
Betty - Beta-hydraoxy butyryl CoA dehydrogenase<br><br />
Deisel Blaze - Butyraldehyde dehydrogenase<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
Meetings (Agenda's, Minutes, Action Items):<br />
<br />
[[Alberta/Calender/Meetings|Meeting Information]]<br />
<br />
<br />
Online Form:<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-27T04:39:05Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichiometric air to fuel ratio at 4 different angular speeds. The experimental setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomass into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum'' (i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli'''s tolerance to solvents such as butanol.<br />
<br />
I: Transforming genes in ''C. acetobutylicum'''s butanoate pathway into ''E.coli''<br><br />
Genes encoding the enzymes in the ''C. acetobutylicum'' butanoate metabolism pathway were identified using the KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy is to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
After receiving the commercially synthesized coding sequences of the genes of interest, the coding sequences are restricted with proper BioBrick prefixes and suffixes out of the original plasmid and cloned into B0034 (ribosome binding site). Double digest (Xba I and Spe I) as well as automated sequencing reactions were performed to verify the proper insertion of the coding sequences and the presence of a ribosome binding site at the 5' end of each coding sequence. <br><br />
<br />
The operon, shown below, consists of all of the genes in the pathway (except ''E. coli'''sthiolase). Note that we purposedly added a Histamine tag to the carboxy terminus of each gene product such that we could use Nickel-NTA column to purify the individual proteins and analyze protein expression and activity.<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
II: Develop Butanol Tolerance with mutagenic and toxic compound Ethylnitrosourea ENU.<br><br />
The concept is to make butanol agar plates of various butanol concentrations and place an ENU disk in the centre. We expect there will be two kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the outer ring of the butanol plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cells that would be mutated in a benifical way to allow them to survive in the butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered three factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''. <br><br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - Butyryl CoA dehydrogenase<br><br />
Enny - Enoyl CoA hydratase<br><br />
Buddy - Butanol dehydrogenase<br><br />
Betty - Beta-hydraoxy butyryl CoA dehydrogenase<br><br />
Deisel Blaze - Butyraldehyde dehydrogenase<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
Meetings (Agenda's, Minutes, Action Items):<br />
<br />
[[Alberta/Calender/Meetings|Meeting Information]]<br />
<br />
<br />
Online Form:<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-26T20:50:02Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichometric air to fuel ratio at 4 different angular speeds. The experimental Setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum''(i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli''s tolerance to solvents such as butanol.<br />
<br />
I: Transforming genes in ''C. acetobutylicum'' butanoate pathway into ''E.coli''<br><br />
Genes encoding the enzymes in the ''C.acetobutylicum'' butanoate metabolism pathway are identified using KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy is to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
After receiving the commercially synthesized coding sequences of the genes of interest, the coding sequences are restricted with proper BioBrick prefices and suffices out of the original plasmid and cloned into B0034 (ribosome binding site). Double digest (Xba I and Spe I) as well as automated sequencing reactions are performed to verify the proper insertion of the coding sequences and the presence of ribosome binding site at the 5' end of each coding sequence. <br><br />
<br />
The operon, shown below, consists of all of the genes of the pathway (except thiolase). Note that we purposedly added a Histamine tag to the carboxy terminus of each gene product such that we could use Nickel-NTA column to purify the individual proteins and analyze protein expression and activity.<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
Objective 2: Develop Butanol Tolerance with mutagenic and toxic compound Ethylnitrosourea ENU.<br><br />
The concept is to make butanol agar plates of various butanol concentrations and place and ENU disk in the centre. We expect there will be two kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the outer ring of the butanol plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cells that would be mutated in a benifical way to allow them to survive in the butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered two factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''.<br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - <br><br />
Enny - <br><br />
Buddy - <br><br />
Betty - <br><br />
Deisel Blaze - <br><br />
J61003 - <br><br />
B0034 - <br><br />
I0500 - <br><br />
Thiolase - <br><br />
BBDBE - Combination of genes in order of ...<br><br />
BEDBB - Combination of genes in order of ...<br><br />
DBEBB - Combination of genes in order of ...<br><br />
DBBBE - Combination of genes in order of ...<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-26T20:49:24Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichometric air to fuel ratio at 4 different angular speeds. The experimental Setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum''(i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli''s tolerance to solvents such as butanol.<br />
<br />
<b> Objective I: Transforming genes in ''C. acetobutylicum'' butanoate pathway into ''E.coli''</b><br><br />
Genes encoding the enzymes in the ''C.acetobutylicum'' butanoate metabolism pathway are identified using KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy is to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
After receiving the commercially synthesized coding sequences of the genes of interest, the coding sequences are restricted with proper BioBrick prefices and suffices out of the original plasmid and cloned into B0034 (ribosome binding site). Double digest (Xba I and Spe I) as well as automated sequencing reactions are performed to verify the proper insertion of the coding sequences and the presence of ribosome binding site at the 5' end of each coding sequence. <br><br />
<br />
The operon, shown below, consists of all of the genes of the pathway (except thiolase). Note that we purposedly added a Histamine tag to the carboxy terminus of each gene product such that we could use Nickel-NTA column to purify the individual proteins and analyze protein expression and activity.<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
Objective 2: Develop Butanol Tolerance with mutagenic and toxic compound Ethylnitrosourea ENU.<br><br />
The concept is to make butanol agar plates of various butanol concentrations and place and ENU disk in the centre. We expect there will be two kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the outer ring of the butanol plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cells that would be mutated in a benifical way to allow them to survive in the butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered two factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''.<br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - <br><br />
Enny - <br><br />
Buddy - <br><br />
Betty - <br><br />
Deisel Blaze - <br><br />
J61003 - <br><br />
B0034 - <br><br />
I0500 - <br><br />
Thiolase - <br><br />
BBDBE - Combination of genes in order of ...<br><br />
BEDBB - Combination of genes in order of ...<br><br />
DBEBB - Combination of genes in order of ...<br><br />
DBBBE - Combination of genes in order of ...<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-26T20:39:01Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichometric air to fuel ratio at 4 different angular speeds. The experimental Setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''Clostridium acetobutylicum''(i.e. endogenous butanoate pathway) into ''E. coli''. Furthermore, we hope to increase ''E. coli''s'' tolerance to solvents such as butanol.<br />
<br />
Objective I: Transforming genes in ''C. acetobutylicum'' butanoate pathway into ''E.coli''<br><br />
Genes encoding the enzymes in the ''C.acetobutylicum'' butanoate metabolism pathway were identified using KEGG database (KEGG number-ca00650: http://www.kegg.com/dbget-bin/www_bget?path:cac00650). Our cloning stragegy was to incorporate all the genes in a single operon with respective inducible promoter and ribosome binding site in a plasmid. Such construct enables easy transformation of multiple genes simultaneously into ''E.coli'' and allows the coordinated expression of genes within the operon. <br><br />
<br />
The operon, shown below, consists of all of the genes of the pathway (except thiolase) and His tags for ease of protien purification.<br />
<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
To develop butanol tolerance with the aid of a mutagen Ethylnitrosourea ENU. The concept is to make butanol agar plates of various butanol concentrations and place and ENU disk in the centre. We expect there will be to kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the ring of the butanol plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cell that were mutated in a benifical way to allow them to survive in the butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered two factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''.<br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - <br><br />
Enny - <br><br />
Buddy - <br><br />
Betty - <br><br />
Deisel Blaze - <br><br />
J61003 - <br><br />
B0034 - <br><br />
I0500 - <br><br />
Thiolase - <br><br />
BBDBE - Combination of genes in order of ...<br><br />
BEDBB - Combination of genes in order of ...<br><br />
DBEBB - Combination of genes in order of ...<br><br />
DBBBE - Combination of genes in order of ...<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-10-26T20:25:04Z<p>CelineYZ: /* '''The Project: Plan B''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns of global greenhouse gasses, the global energy market is in a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. Secondly, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuel source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, organic pollution standards. Blending butanol with gasoline also prevents major modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns as well as offering the possiblity of phase separation which would realize huge cost savings in terms of production.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
<br />
The Butanerds wanted to further evaluate the use of butanol as a fuel in standard spark ignition engines. With the assistance of The University of Alberta's Engine Control Lab in Mechanical Engineering we were able to run a peak power test comparing butanol to iso-octane, a standard test measurement. Each fuel was burned at a stoichometric air to fuel ratio at 4 different angular speeds. The experimental Setup and results can be seen below.<br />
<br />
[[image: alberta_engine1.jpeg|thumb|left|275px| Engine Test Setup Front View]]<br />
[[image: alberta_engine2.jpeg|thumb|left|275px| Engine Test Setup Side View]]<br />
[[image: alberta_enginechart2.jpeg|thumb|left|275px| Peak Power vs. RPM Experimental Data]]<br />
<br><br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''Escherichia coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production from ''''Clostridium acetobutylicum''''(i.e. endogenous butanoate pathway) into ''''E. coli''''. Furthermore, we hope to increase ''''E. coli'''s'' tolerance to solvents such as butanol.<br />
<br />
We have choosen to put all of the genes in a single operon to make the stucture easy to transform as well as so that all genes in the operon have similar expression levels. The operon, shown below, consists of all of the genes of the pathway and his tags for ease of protien purification.<br />
<br />
[[image: alberta_butanol_Operon.jpeg|thumb|center|400px|The Butanol Operon]]<br />
<br />
To develop butanol tolerance with the aid of a mutagen Ethylnitrosourea ENU. The concept is to make butanol agar plates of various butanol concentrations and place and ENU disk in the centre. We expect there will be to kill zones. The first where bacteria die due to the toxicity of ENU at the centre of the plate. The second around the ring of the butanol plate where bacteria die due to butanol toxicity. In between these two zones we hope to see some cell that were mutated in a benifical way to allow them to survive in the butanol.<br />
<br />
[[image: alberta_butanolplate2.jpeg|400px|thumb|center|Proposed Butanol-ENU Plate for Mutagenisis of ''E. coli'']]<br />
<br />
<br />
Concurrently, we are investigating the use of a photoautotrophic bacterium, ''Chlorobium tepidum'' that we will also introduce butanol producing genes to. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as terminal electron donors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees Celsius (104 degrees Fahrenheit), and requires low light conditions for optimal growth. These bacteria grow well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to [[http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html ''here'']]. Images below from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input and output. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
[[image: alberta_carboncycle1.jpeg|thumb|300px|center| Propose Closed Carbon-butanol Cycle]]<br />
<br />
== '''Modeling Efforts''' ==<br />
<br />
Initially our plan to model the efficiency of the butanol production hinged on developing a complete model for the entire glucose pathway in an ''E. coli'' cell using Michaelis-Menten kinetics. After further review, this proved to be beyond the scope of our project's timeline due both to the large number of species and reactions present in the system (as well as their non-linear behavior) and the relative difficulty in finding experimental kinetic information for some of the reactions in our pathway. <br> <br />
<br />
<br />
Therefore, to obtain a rough "first-order" approximation for the behavior of the pathway, we considered two factors:<br />
<br />
1) Stoichiometric factors (both redox and carbon)<br><br />
2) Compared the relative production rates of the various end products of the glucose pathway in ''E. coli'' as found in previous experimentation to the production rates of butanol in ''C. acetobutylicum''.<br />
3) We have also a fuel E. coli Stoichometric Matrix, and intend to stoichiometrically model the system in order to optimize butanol production once the operon is in the system.<br />
<br />
<br />
From these factors we can obtain an indicator of how likely butanol will be produced.<br />
<br><br />
<br />
In order to further develop the system, we hope a full metabolic stochiometric model can be eventually realized for this project. This would allow optimization of the system in order to maximize the flux through the butanol pathway.<br />
<br />
<br />
== '''Project Timeline''' ==<br />
<br />
[[image: Alberta_gantt.jpeg|thumb|400px|center|[[Project Gantt Chart (click here to enlarge)]]]]<br />
<br />
The GANTT Chart Above details the schedule for the lab work that contributed to Plan B. Beginning in July and finishing with our final work in October, the schedule includes details on our different methods to compose an operon that would meet our objectives. In order to read the GANTT Chart for the Lab work of Plan B, please refer to the legend below.<br />
<br />
'''Legend: ''' <br><br />
Benny - <br><br />
Enny - <br><br />
Buddy - <br><br />
Betty - <br><br />
Deisel Blaze - <br><br />
J61003 - <br><br />
B0034 - <br><br />
I0500 - <br><br />
Thiolase - <br><br />
BBDBE - Combination of genes in order of ...<br><br />
BEDBB - Combination of genes in order of ...<br><br />
DBEBB - Combination of genes in order of ...<br><br />
DBBBE - Combination of genes in order of ...<br><br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
[[Alberta/Calender/October|October 2007]]<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team_new.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
== '''Edmonton''' ==<br />
<br />
Welcome to Edmonton!<br />
<br />
<center>[[Image: Alberta_edmonton.jpg]]</center><br />
<br><br />
<br />
For more on the city of Edmonton click '''[http://www.ualberta.ca/~mjl3/About.html here]'''.<br />
<br />
== '''Sponsors''' ==<br />
<br />
We would like to thank all of our sponsers for their gracious support and our advisors for invaluble advise.<br />
<br />
<br />
<b>Major Sponsors</b><br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors1.gif]]<br />
|}<br />
<br />
<br />
'''Sponsors'''<br />
<br />
{| align="center" style="color:white;"<br />
|[[Image: Alberta_sponsors2.gif]]<br />
|}<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta''<br><br />
'''Dr. David Bressler''' - ''University of Alberta''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta''<br><br />
'''Dr. Federick West'''-''University of Alberta''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
'''Dr. David Checkel''' - ''University of Alberta''<br><br />
'''Adrian Audiet''' - ''University of Alberta''<br><br />
'''Dr. Koch''' - ''University of Alberta''<br><br />
'''Dr. Donald Bryant''' - ''Pennsylvania State''<br><br />
'''Dr. Gaozhong Shen''' - ''Pennsylvania State''<br><br />
'''Amaya Garcia''' - ''Pennsylvania State''<br><br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''External Links''' ==<br />
<br />
[http://www.albertaingenuity.ca/ Alberta Ingenuity Fund]<br />
<br />
[http://www.ualberta.ca The University of Alberta Hompage]<br />
<br />
[http://www.mece.ualberta.ca/~ckoch/ Dr. Koch's Engine Lab]<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-10-05T21:36:41Z<p>CelineYZ: /* October 5 */</p>
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font-family: Verdana, Arial, Helvetica, sans-serif;<br />
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<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
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<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
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<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
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<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
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<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
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</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
JG<br><br />
Miniprep J61003+Enny and Benny+J61003<br><br />
Minis are in -20<br><br><br />
ML<br><br />
Brought the tubes labelled "sequencing rxns" up to MBSU<br><br />
Also brought some XBA 1 from fermentas freezer since we ran out<br><br />
Digests JG's miniprep with Xbal and PST. Ran out of out of XBA during digests, which meant that EJ 4,5,6only digested with XBA for 35 min<br><br />
Colony O/N of I0500/ J61003 and Buddy/J61003<br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
VH-1PM<br><br />
No Kan plates therefore made kan plates<br><br />
On COuntertop<br><br />
Miniprepped ML's overnights from OCt 1<br><br />
Lysis solution is a no go<br><br />
Started new O/N of previows overnights for tomorrow<br><br />
No more LB<br><br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
Autoclaved 2 bottosl of Ependorf tupes- to be picked up from G308<br><br />
Transform THolase into XL10 gold plates<br><br />
Miniprep of 10500+J61003 O/N<br><br />
<br />
ML<br><br />
Digest of 10500+J61003 with ECORI and XBA<br><br />
Housekeeping complete<br><br />
Note to Justin: Samples to sequence are in -20 labelled "Justin! Sequence me"<br><br />
CZ - 7:00pm<br><br />
Ran gel of I0500/J61003<br><br />
It looks like I05oo is in J61003 but have to confirm with Justin or Michelle or Erin. <br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
NK - 930<br><br />
VH - 2pm<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs <br><br />
<br />
<br />
CZ - Sorry I can't make it for personal reasons.<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
ED 9:00<br><br />
NK 2pm<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
ED 9:00<br />
<br />
NG 12:00<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 8 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 9 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 10 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 11 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 12 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 13 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 14 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 15 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 16 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 17 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 18 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 19 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 20 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 21 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 22 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 23 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 24 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 25 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 26 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 27 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 28 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 29 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
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== October 30 ==<br />
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[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-10-05T03:00:45Z<p>CelineYZ: /* October 5 */</p>
<hr />
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<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
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<td>M</td><br />
<td>Tu</td><br />
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</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
JG<br><br />
Miniprep J61003+Enny and Benny+J61003<br><br />
Minis are in -20<br><br><br />
ML<br><br />
Brought the tubes labelled "sequencing rxns" up to MBSU<br><br />
Also brought some XBA 1 from fermentas freezer since we ran out<br><br />
Digests JG's miniprep with Xbal and PST. Ran out of out of XBA during digests, which meant that EJ 4,5,6only digested with XBA for 35 min<br><br />
Colony O/N of I0500/ J61003 and Buddy/J61003<br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
VH-1PM<br><br />
No Kan plates therefore made kan plates<br><br />
On COuntertop<br><br />
Miniprepped ML's overnights from OCt 1<br><br />
Lysis solution is a no go<br><br />
Started new O/N of previows overnights for tomorrow<br><br />
No more LB<br><br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
Autoclaved 2 bottosl of Ependorf tupes- to be picked up from G308<br><br />
Transform THolase into XL10 gold plates<br><br />
Miniprep of 10500+J61003 O/N<br><br />
<br />
ML<br><br />
Digest of 10500+J61003 with ECORI and XBA<br><br />
Housekeeping complete<br><br />
Note to Justin: Samples to sequence are in -20 labelled "Justin! Sequence me"<br><br />
CZ - 7:00pm<br><br />
Ran gel of I0500/J61003<br><br />
It looks like I05oo is in J61003 but have to confirm with Justin or Michelle or Erin. <br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
NK - 930<br><br />
VH - 2pm<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs <br><br />
<br />
<br />
CZ - night (should be able to come)<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
ED 9:00<br><br />
NK 2pm<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
ED 9:00<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-10-03T20:18:37Z<p>CelineYZ: /* October 5 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
VH-1PM<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
CZ - 7:00pm<br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
NK - 930<br><br />
VH - 2pm<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs <br><br />
CZ - night (to be confirmed)<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
ED 9:00<br><br />
NK 2pm<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
ED 9:00<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-30T20:26:03Z<p>CelineYZ: /* October 5 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
CZ - 7:00pm<br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs <br><br />
CZ - 2:30pm<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-30T20:25:51Z<p>CelineYZ: /* October 5 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
CZ - 7:00pm<br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs<br />
CZ - 2:30pm<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-30T20:24:16Z<p>CelineYZ: /* October 3 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
CZ - 7:00pm<br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-30T20:23:39Z<p>CelineYZ: /* October 1 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
MC - 800hrs <br><br />
CZ - 2:30pm <br><br />
<br />
<b>NB: please note the lab is unavailable EVERY wednesday from 1400-1700hrs</b><br><br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
MC - 800hrs<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-29T04:57:55Z<p>CelineYZ: /* October 3 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
CZ - 2:30pm<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 == <br><br />
<br />
CZ - 2:30pm<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/OctoberAlberta/Calender/October2007-09-29T04:57:33Z<p>CelineYZ: /* October 1 */</p>
<hr />
<div><div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/October|October 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<br />
<tr><br />
<td></td><br />
<td>[[Alberta/Calender/October#October_1|1]]</td><br />
<td>[[Alberta/Calender/October#October_2|2]]</td><br />
<td>[[Alberta/Calender/October#October_3|3]]</td><br />
<td>[[Alberta/Calender/October#October_4|4]]</td><br />
<td>[[Alberta/Calender/October#October_5|5]]</td><br />
<td>[[Alberta/Calender/October#October_6|6]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_7|7]]</td><br />
<td>[[Alberta/Calender/October#October_8|8]]</td><br />
<td>[[Alberta/Calender/October#October_9|9]]</td><br />
<td>[[Alberta/Calender/October#October_10|10]]</td><br />
<td>[[Alberta/Calender/October#October_11|11]]</td><br />
<td>[[Alberta/Calender/October#October_12|12]]</td><br />
<td>[[Alberta/Calender/October#October_13|13]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_14|14]]</td><br />
<td>[[Alberta/Calender/October#October_15|15]]</td><br />
<td>[[Alberta/Calender/October#October_16|16]]</td><br />
<td>[[Alberta/Calender/October#October_17|17]]</td><br />
<td>[[Alberta/Calender/October#October_18|18]]</td><br />
<td>[[Alberta/Calender/October#October_19|19]]</td><br />
<td>[[Alberta/Calender/October#October_20|20]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_21|21]]</td><br />
<td>[[Alberta/Calender/October#October_22|22]]</td><br />
<td>[[Alberta/Calender/October#October_23|23]]</td><br />
<td>[[Alberta/Calender/October#October_24|24]]</td><br />
<td>[[Alberta/Calender/October#October_25|25]]</td><br />
<td>[[Alberta/Calender/October#October_26|26]]</td><br />
<td>[[Alberta/Calender/October#October_27|27]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/October#October_28|28]]</td><br />
<td>[[Alberta/Calender/October#October_29|29]]</td><br />
<td>[[Alberta/Calender/October#October_30|30]]</td><br />
<td>[[Alberta/Calender/October#October_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/September|September 2007]]<br><br />
To [[Alberta/Calender/November|November 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== October 1 == <br><br />
<br />
CZ - 2:30pm<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 3 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 4 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 5 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 7 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 8 ==<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 9 ==<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 10 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 11 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 12 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 13 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 14 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 15 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 16 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
== October 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/October#October|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/September|To September 2007]]<br><br />
[[Alberta/Calender/Novemeber|To November 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-24T02:31:59Z<p>CelineYZ: /* September 29 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br><br />
Miniprep I0500<br><br />
DB BE and DB-BB transformations did not work.<br><br />
Re-ligated DB into BB boo and DB into BE boo<br><br />
<br />
<br />
11-2:<br />
AL,ML<br><br />
Re-transform BE+DB and BB+DB<br><br />
<br />
2-5:<br />
VH<br><br />
Gel extraction of I0500, but we missed the step "restricting with Ecori+speI" <br><br />
<br />
5-8:<br />
JP,NK<br><br />
Restrction on I0500 mini's with Ecori and SPeI<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br><br />
Double digest of BB-DB/BE-DB XBA/PST<br><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br><br />
Ran gel of the restrictions made in MID, Sept 10 by ML and AL.<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
Restrictions of BB+DB with XBA/PST and BE+DB with PST, SPE<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br><br />
Ran gel on restrictions made by NK in AM of Sept 11<br />
<br />
Night- ED JP<br><br />
Gel extractions<br><br />
Ligations of DB+BB + BE and DB+BE + BB<br><br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH <br><br />
lab class 2-5<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Transformed BEDBB and BBDB into XL 10 gold<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Digested BEBOO and BBBOO with PST and XBA<br />
<br />
PM - VH, JG<br><br />
Ran gel of restrictions made in the previous shift.<br><br />
<br />
Meeting 7:00<br />
<br />
After lab shift, JG, JP<br><br />
Overnights with AMP of BBDB + BEDB<br><br />
Gel extractions from gel made in the prevous shift<br><br />
bands 1,2 (BE) correct while band 3 is too large to be BB<br><br />
Sequence reactions can now be started using primers 1-5 that have been diluted to 5pmol/microlitre in H20 not TE<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL, JG<br><br />
Restrictions on BBDBE and BEDBB with xba/pst and spe/pst<br><br />
PM - VH, CZ<br><br />
Ran gel on restrictions on BBDBE and BEDBB with xba/pst and spe/pst<br><br />
Gel extracions<br><br />
<br />
night - JP<br><br />
Sequencing reactions<br><br />
BBDBBE 6 reactions<br><br />
BEDBBB 6 reactions<br><br />
BB 3 reactions<br><br />
BE 3 reactions<br><br><br />
General note: Each primer sequence will be the end of the indicated gene to show us wether the joining/ligations worked correctly (VF is the promoter/gene/juction)<br><br />
To do: Submit to MBSU<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br><br />
General Note:We have cloned all 5 genes in series, in to differnt orders. Waiting for I0500 which couls be completed on monday. To make sure we have cloned correctly will also need to wait for the squencing results. This weekdn we will clone the 5 gense in 2 differnt orders. The digest have already been done<br><br />
Gel extraction of gel bands of XBA,PST of BBDBBE and BEDBBB. THese can be used to cone into I0500 J61003<br><br />
Ligations of DBBB into BE and DBBE into DBBEBB<br><br />
Leave at 20 degrees celsius for 10 hours<br><br />
5-8 - NK<br />
Cant find the competent cells. <br><br />
Competent cells are in -80 freezer 2nd shelf from the bottom-ML, NG<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br><br />
Transform ligations of DBBEBB and DBBBBE into competent cells. <br><br />
Plated on AMP with whole ligations<br><br />
General Info: When DBBEBB and DBBBBE are complete run sequence reaction<br><br />
Follow "flouresence sequence reaction" protocol<br><br />
<br />
11-2 VH, CZ<br><br />
<br />
2-5 MC, JP<br><br />
<br />
5-8 - ML, NG<br><br />
No growth on DBBBBE or DBBBBE at 1700<br><br />
Retransformed into competent cells DBBEBB and DBBBBE<br><br />
Plated 2 of each and lover overnight at 37 degrees<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs)<br><br />
- delivered MSBU sequencing samples<br><br />
- O/days of Ligations from yesterday<br><br />
<br />
<b>NB: AMP plates are marked with a red streak down the SIDE of the plate. if it does not have this - then it is NOT AMP</b><br><br />
<br />
ML (~10:30-1:00)<br><br />
<br />
<b>To do:</b><br><br />
miniprep of DBBEBB and DBBBBE & glycerol stocks<br><br />
restrict with (1)XBA/PST, (2)SPE/PST<br><br />
Ruun gel of UNCut DBBEBB and DBBBBE, Cut with XBA/PST and CUT with SPE/PST<br><br />
GEl extract XBA/PST bands<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br><br />
Sorry guys, I havent done mini-preps before so just incase I mess up i decided not do it. Instead I made a gel for tonight and update the wiki (even the missing parts from before).<br />
<br />
VH - (@1pm)<br />
<br />
AL - 12-2pm<br />
<br />
NK-6PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
<br />
<b>if you're the first one in; I0500 arrived - pop by and give Mike a visit!:) </b><br />
<br />
MC - in the AM - unsure what time because has to be downtown for a class project @900hrs (hoping to get out of there within a hour) so maybe 10/1030? <br />
<br />
ML - ~10:30 - 1:00<br />
<br />
AL - ~1:00 - 2:00<br />
<br />
Last Wednesday there was a lab class in our lab from 2-5, is this going to be a regular occurence? -VH<br />
<br />
YES - MC <br />
<br />
VH, see notes on Sept 12. - AL<br />
<br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 930 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG, MC- (@5PM)<br />
<br />
JP- late<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs), <br />
<br />
ML (~10:30 - 1:00)<br><br />
<br />
AL - ~12:30 - 2:00<br />
<br />
CZ-2:15PM <br> <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
JP evening<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
Poster Meeting @ 11:00am<br />
Meet in sub By the Subway--JG<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
AM -JG @ 8000hrs <br><br />
MC @ 845 - going to stop by dean of students' to pick up swag first.<br />
<br />
<br />
PM - NK @5 <br><br />
sorry, cant make it till 630<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
930-1230 NK<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
JG & MC - 8:00AM <br><br />
NK 930-1230 <br><br />
CZ 2:30pm<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
CZ 2:00 pm<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-24T02:31:32Z<p>CelineYZ: /* September 28 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br><br />
Miniprep I0500<br><br />
DB BE and DB-BB transformations did not work.<br><br />
Re-ligated DB into BB boo and DB into BE boo<br><br />
<br />
<br />
11-2:<br />
AL,ML<br><br />
Re-transform BE+DB and BB+DB<br><br />
<br />
2-5:<br />
VH<br><br />
Gel extraction of I0500, but we missed the step "restricting with Ecori+speI" <br><br />
<br />
5-8:<br />
JP,NK<br><br />
Restrction on I0500 mini's with Ecori and SPeI<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br><br />
Double digest of BB-DB/BE-DB XBA/PST<br><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br><br />
Ran gel of the restrictions made in MID, Sept 10 by ML and AL.<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
Restrictions of BB+DB with XBA/PST and BE+DB with PST, SPE<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br><br />
Ran gel on restrictions made by NK in AM of Sept 11<br />
<br />
Night- ED JP<br><br />
Gel extractions<br><br />
Ligations of DB+BB + BE and DB+BE + BB<br><br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH <br><br />
lab class 2-5<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Transformed BEDBB and BBDB into XL 10 gold<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Digested BEBOO and BBBOO with PST and XBA<br />
<br />
PM - VH, JG<br><br />
Ran gel of restrictions made in the previous shift.<br><br />
<br />
Meeting 7:00<br />
<br />
After lab shift, JG, JP<br><br />
Overnights with AMP of BBDB + BEDB<br><br />
Gel extractions from gel made in the prevous shift<br><br />
bands 1,2 (BE) correct while band 3 is too large to be BB<br><br />
Sequence reactions can now be started using primers 1-5 that have been diluted to 5pmol/microlitre in H20 not TE<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL, JG<br><br />
Restrictions on BBDBE and BEDBB with xba/pst and spe/pst<br><br />
PM - VH, CZ<br><br />
Ran gel on restrictions on BBDBE and BEDBB with xba/pst and spe/pst<br><br />
Gel extracions<br><br />
<br />
night - JP<br><br />
Sequencing reactions<br><br />
BBDBBE 6 reactions<br><br />
BEDBBB 6 reactions<br><br />
BB 3 reactions<br><br />
BE 3 reactions<br><br><br />
General note: Each primer sequence will be the end of the indicated gene to show us wether the joining/ligations worked correctly (VF is the promoter/gene/juction)<br><br />
To do: Submit to MBSU<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br><br />
General Note:We have cloned all 5 genes in series, in to differnt orders. Waiting for I0500 which couls be completed on monday. To make sure we have cloned correctly will also need to wait for the squencing results. This weekdn we will clone the 5 gense in 2 differnt orders. The digest have already been done<br><br />
Gel extraction of gel bands of XBA,PST of BBDBBE and BEDBBB. THese can be used to cone into I0500 J61003<br><br />
Ligations of DBBB into BE and DBBE into DBBEBB<br><br />
Leave at 20 degrees celsius for 10 hours<br><br />
5-8 - NK<br />
Cant find the competent cells. <br><br />
Competent cells are in -80 freezer 2nd shelf from the bottom-ML, NG<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br><br />
Transform ligations of DBBEBB and DBBBBE into competent cells. <br><br />
Plated on AMP with whole ligations<br><br />
General Info: When DBBEBB and DBBBBE are complete run sequence reaction<br><br />
Follow "flouresence sequence reaction" protocol<br><br />
<br />
11-2 VH, CZ<br><br />
<br />
2-5 MC, JP<br><br />
<br />
5-8 - ML, NG<br><br />
No growth on DBBBBE or DBBBBE at 1700<br><br />
Retransformed into competent cells DBBEBB and DBBBBE<br><br />
Plated 2 of each and lover overnight at 37 degrees<br><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs)<br><br />
- delivered MSBU sequencing samples<br><br />
- O/days of Ligations from yesterday<br><br />
<br />
<b>NB: AMP plates are marked with a red streak down the SIDE of the plate. if it does not have this - then it is NOT AMP</b><br><br />
<br />
ML (~10:30-1:00)<br><br />
<br />
<b>To do:</b><br><br />
miniprep of DBBEBB and DBBBBE & glycerol stocks<br><br />
restrict with (1)XBA/PST, (2)SPE/PST<br><br />
Ruun gel of UNCut DBBEBB and DBBBBE, Cut with XBA/PST and CUT with SPE/PST<br><br />
GEl extract XBA/PST bands<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br><br />
Sorry guys, I havent done mini-preps before so just incase I mess up i decided not do it. Instead I made a gel for tonight and update the wiki (even the missing parts from before).<br />
<br />
VH - (@1pm)<br />
<br />
AL - 12-2pm<br />
<br />
NK-6PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
<br />
<b>if you're the first one in; I0500 arrived - pop by and give Mike a visit!:) </b><br />
<br />
MC - in the AM - unsure what time because has to be downtown for a class project @900hrs (hoping to get out of there within a hour) so maybe 10/1030? <br />
<br />
ML - ~10:30 - 1:00<br />
<br />
AL - ~1:00 - 2:00<br />
<br />
Last Wednesday there was a lab class in our lab from 2-5, is this going to be a regular occurence? -VH<br />
<br />
YES - MC <br />
<br />
VH, see notes on Sept 12. - AL<br />
<br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 930 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG, MC- (@5PM)<br />
<br />
JP- late<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs), <br />
<br />
ML (~10:30 - 1:00)<br><br />
<br />
AL - ~12:30 - 2:00<br />
<br />
CZ-2:15PM <br> <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
JP evening<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
Poster Meeting @ 11:00am<br />
Meet in sub By the Subway--JG<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
AM -JG @ 8000hrs <br><br />
MC @ 845 - going to stop by dean of students' to pick up swag first.<br />
<br />
<br />
PM - NK @5 <br><br />
sorry, cant make it till 630<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
930-1230 NK<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
JG & MC - 8:00AM <br><br />
NK 930-1230 <br><br />
CZ 2:30pm<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-17T18:12:44Z<p>CelineYZ: /* September 19 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br />
<br />
Night- ED JP<br />
<br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH (I will be in at about 1 or 1:30PM, if there is something written down for me to do I can work at it by myself, otherwise if someone wants to come in and get me started on something that would be nice.)<br />
<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Sorry I have meeting at 6pm which should last no longer than 2 hours, so I will come at 8pm the latest.<br><br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Hey guys, i wont be able to make it till 930 AM-NK<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NK, I have it covered.<br />
<br />
Could not do transformation ligations because could not find competent cells. The ligations are in the -20 freezer with the green tape saying "ligations, sept 15". -NK<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs), ML (~10:30-1:00)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br />
<br />
VH - (@1pm)<br />
<br />
AL - 12-2pm<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
ML - ~10:30 - 1:00<br />
<br />
AL - ~12:30 - 2:00<br />
<br />
Last Wednesday there was a lab class in our lab from 2-5, is this going to be a regular occurence? -VH<br />
<br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 8 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG- (@5PM)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs), <br />
<br />
ML (~10:30 - 1:00)<br><br />
<br />
AL - ~12:30 - 2:00<br />
<br />
CZ-2:15PM <br> <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
ED 9-12<br />
<br />
NG 12-3<br />
<br />
NK 230-530<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-17T03:53:33Z<p>CelineYZ: /* September 21 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br />
<br />
Night- ED JP<br />
<br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH (I will be in at about 1 or 1:30PM, if there is something written down for me to do I can work at it by myself, otherwise if someone wants to come in and get me started on something that would be nice.)<br />
<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Sorry I have meeting at 6pm which should last no longer than 2 hours, so I will come at 8pm the latest.<br><br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Hey guys, i wont be able to make it till 930 AM-NK<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NK, I have it covered.<br />
<br />
Could not do transformation ligations because could not find competent cells. The ligations are in the -20 freezer with the green tape saying "ligations, sept 15". -NK<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br />
<br />
VH - (@1pm)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
CZ-2:15PM <br><br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 8 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG- (@5PM)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs)<br><br />
CZ-2:15PM <br> <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
ED 9-12<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-17T03:53:11Z<p>CelineYZ: /* September 19 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br />
<br />
Night- ED JP<br />
<br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH (I will be in at about 1 or 1:30PM, if there is something written down for me to do I can work at it by myself, otherwise if someone wants to come in and get me started on something that would be nice.)<br />
<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Sorry I have meeting at 6pm which should last no longer than 2 hours, so I will come at 8pm the latest.<br><br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Hey guys, i wont be able to make it till 930 AM-NK<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NK, I have it covered.<br />
<br />
Could not do transformation ligations because could not find competent cells. The ligations are in the -20 freezer with the green tape saying "ligations, sept 15". -NK<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br />
<br />
VH - (@1pm)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
CZ-2:15PM <br><br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 8 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG- (@5PM)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs) <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
ED 9-12<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-17T03:52:56Z<p>CelineYZ: /* September 19 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once<br />
digested all in Boos except for DB with Pst/xba<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br><br />
put away bottled LB<br><br />
Ran gel of last night's restricted samples "in boo"; did not really turn out<br><br />
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing<br><br />
Mini prepped Ligations of DB in BB and DB in BE<br><br />
<br />
to do:<br />
- run gel of double digested I0500<br><br />
- do single & double restriction on Mini prep of Ligations<br><br />
- run gel of uncut, single & double restrictions of mini prepped ligations<br><br />
<br />
ML,AL for the MID <b>(AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)</b><br />
<br />
NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
Jason if you have time to show up that would be sweet, but if you don't I understand. -NG<br />
<br />
<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL (12:30 - 2 ish), NG (random)<br />
<br />
Night- ED JP<br />
<br />
<b> mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!</b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
NG <b> Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).</b><br />
<br />
AM - <br />
<br />
Mid- ML, AL<br />
<br />
<b> Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL</b><br />
<br />
PM - VH (I will be in at about 1 or 1:30PM, if there is something written down for me to do I can work at it by myself, otherwise if someone wants to come in and get me started on something that would be nice.)<br />
<br />
<b> VH: please make note of the above announcement regarding lab class at 2:00pm </b><br />
<br />
Night - CZ, ED<br><br />
Sorry I have meeting at 6pm which should last no longer than 2 hours, so I will come at 8pm the latest.<br><br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
Hey guys, i wont be able to make it till 930 AM-NK<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM - MC (around 900/930hrs)<br><br />
- Mini prep from O/N of BBDBE & BEDBB<br><br />
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates<br><br />
- Gel extraction of BE & BB Xba/Pst<br><br />
- made gel<br> <br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NK, I have it covered.<br />
<br />
Could not do transformation ligations because could not find competent cells. The ligations are in the -20 freezer with the green tape saying "ligations, sept 15". -NK<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
AM - JG (800 hrs - 930hrs),MC (@ 800hrs)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
NK - 8 AM<br />
<br />
VH - (@1pm)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
CZ-2:15PM<br />
ED- 6PM <br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
NK - 8 AM<br />
<br />
VH - (@1PM)<br />
<br />
JG- (@5PM)<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
AM - MC, JG (@ 800hrs) <br />
ED - 4 PM<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
ED 9-12<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
ED 9-12<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-10T03:14:48Z<p>CelineYZ: /* September 12 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br />
<br />
ML,AL for the MID<br />
<br />
NG for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL<br />
<br />
Night- ED JP<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
AM - MC <b> Sorry this is one of the "odd" wednesdays i am unable to make it in the AM until 1130hrs. . . Or i can come in from 800hrs to 920ish. . . but then i have go jet for 2hrs</b><br />
<br />
Mid- ML, AL<br />
<br />
PM - VH, NG<br />
<br />
Night - CZ, ED<br><br />
Sorry I have meeting at 6pm which should last no longer than 2 hours, so I will come at 8pm the latest.<br><br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM- MC, JG<br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NG, ( this is what i had copied down but if anyone else can take this im sure nick would be greatful)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-10T03:13:58Z<p>CelineYZ: /* September 9 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
To [[Alberta/Calender/October|October 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br />
<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<b>Schedule:</b><br />
<br />
JP,ML,JG 7:00pm <br><br />
<br />
<br />
<u>For the record: </u><br><br />
Buddy = BuOH DH 1235 bp <br><br />
Betty = Bb-bhBu-coa dh 923 bp <br><br />
Benny = Butyryl coa dh 1184 bp <br><br />
enny = Enoyl coa Hydratase 860 bp <br><br />
Diezel Blaze = buAld-Dh 2639 bp<br><br />
<br />
<b> Notes:</b> <br><br />
Moving was done this mid-day. <br><br />
New location <b>M349</b> <br><br />
JG has key & access <br><br />
- MC, ED, JG <br><br />
<br />
Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC <br><br />
<br />
<b>Night Crew</b><br />
<br />
-JP, JG<br />
<br />
Set up new lab <br />
<br />
Things we need <br />
- Water Bath, Scale, disposable 13ml overnight tubes, Tip waste<br />
<br />
<b> lab work </b><br />
<br />
Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab<br />
<br />
To Do list for tomorrow<br />
<br />
- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)<br><br />
<br />
-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished! <br><br />
<br />
- If I0500 comes in from calgary it needs to be transformed.<br />
<br />
- Transform R0080 which is a back up promoter just in case?<br />
<br />
<br />
<br />
-Note we also have to do sequenceing on BB and BE and DB<br />
<br />
Shout out to calgary for saving our necks <br />
Shout out to the move in crew <br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<b>Schedule:</b><br />
<br />
NG,ED,ML 7:00pm <br><br />
<br />
<b>AM Crew Notes:</b><br><br />
<br />
finished up cleaning up post-move in<br> <br />
got a 'water bath' - JG calibrated it<br><br />
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow<br><br />
Restricted:<br><br />
- DB in BOO with PST/XBA<br><br />
- DB in BOO with PST/SPE<br><br />
- BB in BOO with PST/SPE<br><br />
- BE in BOO with PST/SPE<br><br />
<b>NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC </b> <br><br />
<3 JG & MC <br><br />
<br />
Transformed I0500 from Calgary.<br />
Transformed R0080.<br />
Ran digestions on gel.<br />
Ligated digested DB with BB and BE. (all in Boo)<br />
<br />
-ED, ML<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<b>Schedule:</b><br />
<br />
NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)<br><br />
<br />
<b>To Do:</b><br><br />
Ligations<br><br />
<br />
<B> Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up! </b><br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 7 ==<br />
<b>Schedule:</b><br />
<br />
AM Crew: MC, JG <b>meeting at 800hrs</b><br><br />
<br />
<b>Lab Notes:</b><br><br />
No growth observed for R0080 overnights; just scrapping it because we have I0500<br><br />
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details<br><br />
O/N with AMP for I0500<br><br />
Transform religations in to XL10 Gold<br><br />
-MC, JG, ML<br />
<br />
<b>NB: JG has key access</b><br><br />
<br />
<b>To Do:</b><br><br />
Mini preps on I0500 overnights<br><br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
<b>Schedule:</b><br />
<br />
Super Awesome Marathon of Productiveness<br />
<br />
8-11:<br />
JG,NG<br />
<br />
11-2:<br />
AL,ML<br />
<br />
2-5:<br />
VH,NK<br />
<br />
5-8:<br />
JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
<b>Schedule:</b><br />
<br />
And Yet another super productive Sunday<br />
<br />
8-11: NG and CZ<br />
<br />
NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.<br />
<br />
NG - Update: 9:30am 'broke' into lab :D. Then found key...<br />
<br />
11-2: VH, CZ<br><br />
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb). <br><br />
Digested I0500 and J61003 with Eco/Pst as instructed.<br><br />
<br />
2-5: MC, JG<br><br />
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either<br><br />
gel extracted J61003<br><br />
Made LB - to be divided into bottles & autoclaved in G308<br><br />
O/N of DB in BB & DB in BE samples left in 37degree room<br><br />
<br />
5-8: ML, ED<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
<b>Schedule:</b><br />
And Scheduling just got hard core<br />
<br />
I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could.<br />
-JG<br />
<br />
MC,JG for the AM<br />
<br />
ML,AL for the MID<br />
<br />
NG for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG)<br />
<br />
JP,ED for the Night<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
<br />
<b>Schedule:</b><br />
No one else really signed up for tuesdays so anyone who has time to drop in should<br />
<br />
AM- NK<br />
<br />
Mid - AL<br />
<br />
Night- ED JP<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
<b>Schedule:</b><br />
<br />
AM - MC <b> Sorry this is one of the "odd" wednesdays i am unable to make it in the AM until 1130hrs. . . Or i can come in from 800hrs to 920ish. . . but then i have go jet for 2hrs</b><br />
<br />
Mid- ML, AL<br />
<br />
PM - VH, NG<br />
<br />
Night - CZ, ED<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
<b>Schedule:</b><br />
<br />
AM-Nik<br />
<br />
PM - VH, JG<br />
<br />
Meeting 7:00<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AM- MC, JG<br />
<br />
Mid - AL<br />
<br />
PM - VH, CZ<br />
<br />
night - JP<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
<b>Schedule:</b><br />
<br />
And yet another crazy weekend...<br />
<br />
8-11 - ED,MC<br />
<br />
11-2 - JG, ML<br />
<br />
2-5 - NG, CZ<br />
<br />
5-8 - NG, ( this is what i had copied down but if anyone else can take this im sure nick would be greatful)<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
<b>Schedule:</b><br />
Continued...<br />
<br />
8-11 ED, JG<br />
<br />
11-2 VH, CZ<br />
<br />
2-5 MC, JP<br />
<br />
5-8 - ML, NG<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
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<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 25 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 26 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 27 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 28 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 29 ==<br />
<br />
<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-01T19:41:01Z<p>CelineYZ: /* '''September''' */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br><br />
<br />
<b> Lab Notes </b><br><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 3 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 4 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 6 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
== September 7 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 8 ==<br />
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<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 9 ==<br />
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<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 10 ==<br />
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<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 11 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 12 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 13 ==<br />
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<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 14 ==<br />
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<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 15 ==<br />
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<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 16 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 17 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 18 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 19 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 20 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 21 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 22 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 23 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 24 ==<br />
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<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
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== September 26 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 27 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 28 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 29 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 30 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/SeptemberAlberta/Calender/September2007-09-01T19:40:30Z<p>CelineYZ: /* September 1 */</p>
<hr />
<div>=='''September'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/September|September 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/September#September_1|1]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_2|2]]</td><br />
<td>[[Alberta/Calender/September#September_3|3]]</td><br />
<td>[[Alberta/Calender/September#September_4|4]]</td><br />
<td>[[Alberta/Calender/September#September_5|5]]</td><br />
<td>[[Alberta/Calender/September#September_6|6]]</td><br />
<td>[[Alberta/Calender/September#September_7|7]]</td><br />
<td>[[Alberta/Calender/September#September_8|8]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_9|9]]</td><br />
<td>[[Alberta/Calender/September#September_10|10]]</td><br />
<td>[[Alberta/Calender/September#September_11|11]]</td><br />
<td>[[Alberta/Calender/September#September_12|12]]</td><br />
<td>[[Alberta/Calender/September#September_13|13]]</td><br />
<td>[[Alberta/Calender/September#September_14|14]]</td><br />
<td>[[Alberta/Calender/September#September_15|15]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_16|16]]</td><br />
<td>[[Alberta/Calender/September#September_17|17]]</td><br />
<td>[[Alberta/Calender/September#September_18|18]]</td><br />
<td>[[Alberta/Calender/September#September_19|19]]</td><br />
<td>[[Alberta/Calender/September#September_20|20]]</td><br />
<td>[[Alberta/Calender/September#September_21|21]]</td><br />
<td>[[Alberta/Calender/September#September_22|22]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_23|23]]</td><br />
<td>[[Alberta/Calender/September#September_24|24]]</td><br />
<td>[[Alberta/Calender/September#September_25|25]]</td><br />
<td>[[Alberta/Calender/September#September_26|26]]</td><br />
<td>[[Alberta/Calender/September#September_27|27]]</td><br />
<td>[[Alberta/Calender/September#September_28|28]]</td><br />
<td>[[Alberta/Calender/September#September_29|29]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/September#September_30|30]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
</tr><br />
<br />
<br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== September 1 ==<br><br />
<br />
<b> Lab Notes </b><br />
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer<br><br />
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights<br><br />
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later<br><br />
<br />
-CZ<br />
<br />
<br />
<br />
[[Alberta/Calender/September#September|to the top]]<br />
<br />
== September 2 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 3 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 4 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 5 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 6 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 7 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 8 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 9 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 10 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 11 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 12 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 13 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 14 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 15 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 16 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 17 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 18 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 19 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 20 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 21 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 22 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 23 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 24 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 25 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 26 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 27 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 28 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 29 ==<br />
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[[Alberta/Calender/September#September|to the top]]<br />
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== September 30 ==<br />
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<br />
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[[Alberta/Calender/September#September|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/August|To August 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-09-01T03:15:15Z<p>CelineYZ: /* August 31 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK <br><br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store at the front counter overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK <br><br />
hey guys, iv got a meeting at 6:30, so i might be a little late-NK<br />
<br />
<b>General Notes:</b><br />
<br />
1. after comparing the sequenced Diezel Blaze samples (1,2,4,7) with Erin's sent out sequences, the construct is not correct. Therefore tonight we must complete restriction of D.B with XbaI/PstI and re-ligate it in Boo that has been restricted with XbaI/PstI.<br />
<br />
<b>Lab Notes</b><br><br />
1. After talking to James, we decided to do a triple digest (Pst/Xba/SspI). SspI is an unique site inside the D.B. vector (i.e.pUC57) but absent inside D.B. Ssp works in Buffer O. So Pst/SspI digest in Buffer O for 1.5 hours at 37 degree, then Xba in Buffer Tango for 1 hour. Also we did a negative control where D.B. is digested with SspI only. <br> Both tubes are stored in a box with white tape in the -20 freezer. Gel is already pour for tomorrow.<br><br />
2. Transformation of yesterday's ligation (Buddy+Benny, Betty+Enny) into DH5alpha and stored in 37 degree incubator overnight.<br> <br />
3. Appears to be some <b>Chlorobium growth</b> - MEDIUM WORKS! - the next step is to make plates and see if they work - making the plates will happen on thursday<br><br />
<b>For Tomorrow</b><br />
1. Run gel of tonight's digest. For the negative control, one should expect a band around 5.3kb(unique site, linearized vector 2.7kb + D.B. 2.6kb). For the triple digest, one should expect the D.B. band at 2.7kb and the vector band at 2.0kb. The upper 2.7 kb is the one to be cut out, gel purified, and ligate with Boo like previously overnight.<br><br />
2. If there are colonies from tonight's transformation, colony PCR and run gel if they are ligated together properly.<br><br />
In attendance:<br><br />
JP, CZ, NK <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
I will drop by briefly to see the gels and the colonies out of curiosity. -CZ,JP<br><br />
AL,JG<br />
<br />
<b>Also To Do List (in addition to those listed yesterday): </b><br><br />
<br />
1- restart I0500 PCR + run all 50microL in gel to purify maximum amount -<b>we need this to work because Meagan at MIT hasn't emailed back!</b> <br><br />
<br />
<b> Lab Notes </b><br />
<br />
Ran gel on previous nights digest, 2 triple digest and a -ve control. We also Colony PCR'ed buddy and boo + benny boo and Buddy boo + enny boo as well as 2 I0500. Cut and purified gel of D.B. and then ligated with boo. left on front bench.<br />
<br />
<br />
NOTE: PCR was left on the machine.<br />
<br />
Quote:<br />
"DAMN We are Goood!!!"<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG,NK <br><br />
ligations from previous night were not what was expected on gel therefore re-ligated Diezel blaze (xba/pst) with B0034 (speI/Pst. The ligations left at front bench. <br><br />
ran gel of colony PCR products. Only one of the picked colonies (betty and Enny in boo) had correctly sized insert. The colony could not be matched to the sample, so we started overnights of 4 betty and Enny in boos and 8 benny and buddy in boos.<br><br />
<b>To do next lab:</b><br><br />
1)Mini-prep of buddy-benny in boo and betty-enny in boo <br><br />
2)Restrict 4microL of the mini'd overnights with Xba/Pst - run sample in gel to determine correct insert <br><br />
3)Transformation of the re-lighated Diezel blaze in B0034<br><br />
4)PCR of I0500 again. This time use following recipe (as we will need lots!)<br><br />
<br />
-40.5micoL water<br><br />
-0.5microL taq<br><br />
-5.0microL 10X buffer<br><br />
-0.75microL of each primer<br><br />
-2.5microL of I0500 template<br><br />
<br />
1- Run @ 95 C for 2:00<br><br />
2- Run @ 95 C for 0:30<br><br />
3- Run @ 60 C? for 0:30 (check Tm of primers and go anneal = Tm - 5 C)<br><br />
4- Run @ 72 C for 2:00<br><br />
***repeat 2-4 30 times***<br><br />
5- Run @ 72 C for 5:00<br><br />
<br />
5)WE NEED MORE DNA LADDER!<br><br />
6)Make chlorobium plates <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK, NG<br><br />
NK and JG are meeting up at 3:00 PM, whoever else wants to meet up can meet up then-NK<br><br><br />
<b>MC available in the AM - because she has to make some phone calls to peoples from the lab - please let me know if you need any work done that can be done within 3hrs via email</b> <br><br />
<br />
<b>AM Lab Notes:</b><br><br />
Miniprepped all 12 samples; in -20freezer<br><br />
Transformed religated DB in B0034 into XL10Gold and plated 200ul (x5) of each - left in 37 incubator <br><br />
<br />
-MC<br />
<br />
<b>To Do List:</b> <br><br />
<br />
1 - make Chlorobium plates with and without antibiotics and put them in Capital Health anaerobic chamber<br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1-completed mini's of Buddy and Benny in boo and Betty and enny in boo<br><br />
2-completed restrictions (Xba/Pst) of mini'd samples<br><br />
3-ran PCR of I0500 <br><br />
4-ran gel of restricted products and PCR of I0500 - no I0500 - bands for BE and BB may be correct, for better resolution, we ran for longer period after we left last night - NEEDS TO BE CHECKED on friday<br><br />
5-made Chlorobium plates - 7 amp/7 non-selective and they are in anaerobic chamber (capital health)<br><br />
<br />
-MC,JG,ML,AL,JP, DOUG, NK<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br><br />
The second gel on the night of aug 30 was not what was expected either. Our Conclusion: The digest likely did not work. However upon Jame's examination of the gel, BB6,7,8 did seem to be right so those bands were cut out and purified for later use.<br><br />
<br />
Therefore we took the following steps:<br><br />
1. Re-digest mini'd samples->Benny+Buddy Pst/Xbal and Betty+Enny PST/XBal; after running the gel, Betty+Enny4 seemed to work so the band was cut out. It needs to be purified tomorrow.<br><br />
2.Re-ligate: buddy (xbal/pst)+Benny(spe/PST) and Betty(xbal/PST)+Enny(spe/PST); then transformation of these ligation mixtures<br><br />
3.Re-transform previousligation mixture :Buddy/enny+Betty/Enny<br><br />
4. Overnights for Diezel blaze (4ml LB+4microlitreAMP per colony); ready for miniprep tomorrow<br><br />
Also Note:<br><br />
We are now using a new batch of XL10 gold competent cells so transformations with Ing pBS(bluescript)as positive control. Same procesdure as before except for our plasmids plate out 200microlitere, but for the positive control, 2 plates of 100 microlitre and 2 plates of 20 microlitre.<br><br />
<br />
<b> For tomorrow, CZ and ML will meet up between 9:30 and 10:00 to do miniprep for the D.B. as well as gel purification of the Betty+ Enny 4 Pst/Xba digest. However, we are not sure if we will have enough time to pick colonies for the Benny+Buddy and Enny+Betty. If anyone could come in to do that, contact either the lab or CZ. </b> <br><br />
<br />
Attendence-NK, CZ, ML, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-28T03:13:24Z<p>CelineYZ: /* August 28 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK <br><br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store at the front counter overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK <br><br />
hey guys, iv got a meeting at 6:30, so i might be a little late-NK<br />
<br />
<b>General Notes:</b><br />
<br />
1. after comparing the sequenced Diezel Blaze samples (1,2,4,7) with Erin's sent out sequences, the construct is not correct. Therefore tonight we must complete restriction of D.B with XbaI/PstI and re-ligate it in Boo that has been restricted with XbaI/PstI.<br />
<br />
<b>Lab Notes</b><br />
1. After talking to James, we decided to do a triple digest (Pst/Xba/SspI). SspI is an unique site inside the D.B. vector (i.e.pUC57) but absent inside D.B. Ssp works in Buffer O. So Pst/SspI digest in Buffer O for 1.5 hours at 37 degree, then Xba in Buffer Tango for 1 hour. Also we did a negative control where D.B. is digested with SspI only. <br> Both tubes are stored in a box with white tape in the -20 freezer. Gel is already pour for tomorrow.<br><br />
2. Transformation of yesterday's ligation (Buddy+Benny, Betty+Enny) into DH5alpha and stored in 37 degree incubator overnight.<br> <br />
<b>For Tomorrow</b><br />
1. Run gel of tonight's digest. For the negative control, one should expect a band around 5.3kb(unique site, linearized vector 2.7kb + D.B. 2.6kb). For the triple digest, one should expect the D.B. band at 2.7kb and the vector band at 2.0kb. The upper 2.7 kb is the one to be cut out, gel purified, and ligate with Boo like previously overnight.<br><br />
2. If there are colonies from tonight's transformation, colony PCR and run gel if they are ligated together properly.<br><br />
In attendance:<br><br />
JP, CZ, NK<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
I will drop briefly to see the gels and the colonies out of curiosity. -CZ<br><br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-28T02:36:12Z<p>CelineYZ: /* August 27 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK <br><br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store at the front counter overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK <br><br />
hey guys, iv got a meeting at 6:30, so i might be a little late-NK<br />
<br />
<b>General Notes:</b><br />
<br />
1. after comparing the sequenced Diezel Blaze samples (1,2,4,7) with Erin's sent out sequences, the construct is not correct. Therefore tonight we must complete restriction of D.B with XbaI/PstI and re-ligate it in Boo that has been restricted with XbaI/PstI.<br />
<br />
<b>Lab Notes</b><br />
1. After talking to James, we decided to do a triple digest (Pst/Xba/SspI). SspI is an unique site inside the D.B. vector (i.e.pUC57) but absent inside D.B. Ssp works in Buffer O. So Pst/SspI digest in Buffer O for 1.5 hours at 37 degree, then Xba in Buffer Tango for 1 hour. Also we did a negative control where D.B. is digested with SspI only. <br> Both tubes are stored in a box with white tape in the -20 freezer. Gel is already pour for tomorrow.<br><br />
2. Transformation of yesterday's ligation (Buddy+Benny, Betty+Enny) into DH5alpha and stored in 37 degree incubator overnight.<br> <br />
<b>For Tomorrow</b><br />
1. Run gel of tonight's digest. For the negative control, one should expect a band around 5.3kb(unique site, linearized vector 2.7kb + D.B. 2.6kb). For the triple digest, one should expect the D.B. band at 2.7kb and the vector band at 2.0kb. The upper 2.7 kb is the one to be cut out, gel purified, and ligate with Boo like previously overnight.<br><br />
2. If there are colonies from tonight's transformation, colony PCR and run gel if they are ligated together properly.<br><br />
In attendance:<br><br />
JP, CZ, NK<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-26T19:19:02Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK <br><br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store at the front counter overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-26T18:44:22Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK <br><br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store in 15 degree incubator overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-26T18:44:07Z<p>CelineYZ: /* August 25 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on <br><br />
<br />
In attendance: NK, MC, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store in 15 degree incubator overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-26T18:43:33Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<b> Lab work </b> <br><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store in 15 degree incubator overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-26T18:43:16Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<b> Lab work </b><br />
1. Gel extraction; labelled as<br />
Benny in Boo (1) pst/spe gel purified <br><br />
Benny in Boo (3) pst/spe gel purified <br><br />
Enny in Boo (6) pst/spe gel purified <br><br />
Enny in Boo (9) pst/spe gel purified <br><br />
<br />
2. ligation <br><br />
a. Buddy in Boo pst/xba + Benny in Boo (1) pst/spe <br><br />
b. Buddy in Boo pst/xba + Benny in Boo (3) pst/spe <br><br />
c. Betty in Boo pst/xba + Enny in Boo (6) pst/spe <br><br />
d. Betty in Boo pst/xba + Enny in Boo (9) pst/spe <br><br />
<br />
Store in 15 degree incubator overnight for transformation tomorrow. <br><br />
<br />
In attendance: AL, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-25T22:24:06Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC <br><br />
If you are having trouble mangling gels, you should carry them to the camera inside of the tray that you pour them in. <br><br><br />
<br />
Lab Notes<br><br />
The I0050 sample on gel from august 24th is not new, it is the PCR sample<br><br />
It did not show on the gel <br><br />
Who purified Buddy and betty?<br><br><br />
Lab work<br><br />
PST/Spe restriction of Benny+boo (1,3)<br><br />
PST/Spe restriction of Enny+boo (6,9)<br><br />
Used 4 microlitre of sample<br><br />
Ran gel at 100V<br><br />
Dishwashed and LAB clean up, ethanoled all benches we usually work on<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<b> Meet at noon as usual -CZ </b><br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-25T04:48:49Z<p>CelineYZ: /* August 26 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
2-<b>We need to be making better lab notes in the notebook </b>-<br />
<br />
"Started Overnights of..." is not acceptable. Instead, write what you added to the tube: 5uL Amp, 5mL LB, colony.<br />
<br />
Likewise, "Double digest" is not acceptable. You need to write down what you digested and what amounts of everything has been added.<br />
<br />
Writing in a notebook correctly is very important with this many people in the lab. If people are forgetting to add buffer to digest or antibiotics to overnights, this needs to be corrected immediately and by reading what was done in the lab book, it can be. '''It is much better to write too much than too little, especially if it can jeopardize the future of the project.'''<br />
<br />
I know that I have been guilty of this, but we all need to improve. '''Everything needs to be written down.'''<br />
<br />
<br />
'''Lab Notes:'''<br />
<br />
-I took out plates for I13453 this morning and there are big colonies to pick tonight. <br><br />
-Started overdays of Diezel Blaze and I13453. <br><br />
-Digested D.B with Xba and Pst. <br><br />
-Poured gel for use tonight. <br><br />
<br />
-MC, ED<br />
<br />
Tonight: <br />
<br />
-Miniprep and digest I13453 with EcoR1 and Xba tonight. Miniprep and store D.B in freezer tonight (digest with XbaI/PstI). <br><br />
-made glycerol stocks of each D.B and I13453 <br><br />
<br />
<br />
<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<b>To Do: </b><br />
<br />
1-run gel of D.B restriction products <br><br />
2-gel isolate D.B (XbaI/PstI) <br><br />
3-start ligation of D.B in Boo <br><br />
<br />
<b>Things We're Feeling - a monologue by Justin & Michelle (& Matt - he sends his apologies):</b><br><br />
<br />
Ran a gel for 2 hours to try and resolve D.B (2650bp) and pUC17 (2700bp); don't laugh<br><br />
Diezel Blaze is actually kind of the same size as the plasmid it came in (PUC17)<br><br />
<br />
there were tears<br><br />
<br />
some precision cutting happened - we're going to chance it <br><br />
we hope that most of what was cut out will have mostly Diezel Blaze in it - however to compensate when Ligations are done B0034(vector) should be added in EXCESS to increase chances of B0034 + Diezel Blaze = <3 <br><br />
<br />
we'll have to do some colony PCR with VF/VR after transformation and the rxn's that work will be BOO34 and the ones with inserts the right size will be the correct configuration. We'll definately need to sequence... <br><br />
<br />
end scene<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- gel extraction of D.B (in new box in -20)<br><br />
- Ligation with excess B0034 ( 2microL Boo + 2microL D.B. and 2microL Boo + 4microL D.B.)<br><br />
- expecting bacterial stub of I0050 in the mail if it is in - then grow it up<br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<b>Lab notes:</b><br />
<br />
Gel extract Diezel Blaze using QIAgen Kit.<br />
<br />
Ligated product with BOO34. (1:2 and 1:1 vector: insert ratio)<br />
<br />
Also James dropped by and was wondering about sequencing. The sequencing reaction protocol is at the front counter and takes about 2 hours, perhaps we can do it on this weekend for all of the plasmids we have so far.<br />
<br />
Where are all the plasmid maps for the genes just in case we need to find it?<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
I can go in before. - ED<br />
<br />
What time are we meeting up for the lab? -AL<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed ligations into XL10s and plated on Amp. Plated 50uL and then spun down the remainder of the tube and plated that.<br />
<br />
Tomorrow:<br />
<br />
Colony PCR of Diezel Blaze in Boo and find out what is going on with I0500.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>NB: Meeting for ANAEROBIC LAB tour at 1450hrs at the University of Alberta Hospital "starbucks" across from the gift shop</b><BR><br />
<br />
<b>Schedule:</b> <br><br />
<br />
ED, AF, JG <br><br />
<br />
<b> Morning Lab Party:</b><br><br />
<br />
Sorry Jason - I did not see you were available until now . . . . . <br><br />
Colony PCR (14 samples) from the Diezel Blaze in B0034 transformations <br><br />
- PCR samples will be in the left Thermocycler<br><br />
- Restreaked plates of PCR samples in the 37 degree incubator <br><br />
- Put the transformation plates in the 4 degree fridge in the back without parafilm<br><br />
-MC<br />
<br />
'''Evening Lab Notes:'''<br />
<br />
Poured Amp plates.<br><br />
Digested BuddyB BennyB BettyB EnnyB (B=Boo) with Xba.<br><br />
Ran gel of colony PCRs of Diezel Blaze in Boo.<br> Got two differend band sizes, one at about 2700 and one about 3600. <br> Will grow up both so they can be submitted for sequencing <br><br />
<br />
-ED,CZ,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
Lab marathon this weekend! Start @ 10:00 and goes all day. Everyone show up. We will be leaving mid-afternoon and returning at 8:00 PM.<br />
<br />
I'll be there at 1100ish - AL, jp<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK <br><br />
<br />
<b> Lab Notes:</b><br><br />
Continued with digest from Aug17 & Ran gel - results were different from expected <br><br />
Overdays of Diezel Blaze in Boo34<br><br />
Repeated Digests & ran gel again - unfortunately results were different from expected AND previous gel's results<br><br />
Glycerol stocks of Diezel Blaze in B0034<br><br />
Mini Prep Diezel Blaze in Boo34 <br><br />
<br />
- MC, ED, JP, AL, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Lab Marathon Continues</b><br />
<br />
Meet at noon for more marathon goodness.<br />
<br />
Oops on FAVA!!!!!<br />
They have given away the camera the Anthony had reserved so there will be no movie this weekend :(<br><br />
<br />
<b>Lab Notes:</b><br><br />
Sequencing of all genes (BennyB, BuddyB, BettyB, DiezelBlazeB, EnnyB) in B0034 - only did two of each in the case where there were more than two mini preps<br><br />
stored at -20freezer taped together labled "to MSBU" - will be delivered to MSBU tomorrow<br><br />
<br />
-MC, JG, CZ<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<b>Schedule:</b><br />
<br />
ED,JP,JG<br />
<br />
Digested all "in Boos" for further confirmations, but failed to restrict DB #2 and #7.<br />
<br />
Prepared chemicals for anaerobic broth.<br />
<br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers men, bake me a cake as fast as you can"<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<b>Schedule:</b><br />
<br />
AF,JP,ED <br><br />
<br />
<br />
<b> Lab Notes: </b><br />
<br />
1-ran gels of re-digested "in Boo's" for further confirmation. Realized that D.B samples 2 + 7 should have been restricted too! <br><br />
2-confirmed sequencing results - Buddy and D.B. need to be re sequenced - D.B. 2+7 along with Buddy samples (started overnites tonight - mini tomorrow - digest/sequence) All others are grandioso <br><br />
<br />
3-Started overnights of Buddy in boos which will need to be restricted with Eco/speI (run for size confirmation -but KEEP nicely labeled samples for isolation of RBS-Buddy for puting in J61003) <br><br />
<br />
4-email Meagan from MIT again to see where our bacteria is! <br><br />
<br />
5- make trace elements for Chlorobium culture <br><br />
<br />
<b>Childrens Story of the Day</b> <br><br />
<br />
"Paddy cake, paddy cake, bakers (WO-)men, bake me a cake as fast as you can"<br />
<br />
-ED, JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<b>Schedule:</b><br />
<br />
ED, AL, MC<br />
<br />
<b>General Notes:</b><br />
<br />
1- Dr. Foght was able to give us 25ml of 0.1mg/mL Resazurin she also gave us CuCl2 2H20, FeCl2 4H20, NaSe03 (to replace NaHSeO3- careful its poisonous!), VoSO4 2H20!!! We got more than we barained for!!! Thanks Dr. Foght! - jp<br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprepped Buddy overnight (Buddy in B0034), stored in -20 degree freezer.<br />
<br />
Sequencing prep for Diezel Blaze #2&7. <br />
<br />
PCR I0500, ran on colony PCR in right thermocycle. (Need to be run on gel tomorrow, in PCR machine on the right.)<br />
<br />
Digested all with Pst; Buddy and Betty with Xba; Benny and Enny with Spe. Samples put in freezer, to be run on gel tomorrow.<br />
<br />
Updated cloning progress in lab book. <br />
<br />
2 gels poured for tomorrow. <br />
<br />
Trace elements, salt A, B and C for the Clostridium made: dissolved in distilled water and autoclaved. Left on bench to be cooled. '''Trace elements are at 0.5 X''''<br />
<br />
-ED, MC, AL, AF<br />
<br />
Friendly tip of the day:<br />
"Always wear gloves when handling EtBr! "<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Performed Gel Extraction on I0500 gel August 23rd AM<br />
<br />
Cut out bands for Buddy restricted with Xba and Pst and Betty restricted with Xba and Pst from August 23rd AM gel 2. <br />
<br />
The Benny and Enny restrictions DID NOT work.<br />
<br />
Chlorobium medium is complete and ready for innoculation at 1700 August 24th.<br />
<br />
Emailed Meagan at MIT to ask for I0500 again (incase our PCR'ing doesn't work) <br><br />
<br />
In attendance:<br />
<br />
ML, JP, AF, JG, AL<br />
<br />
<b>Things for tommorow/weekend:</b><br />
<br />
1- <b>Restrict I0500 samples</b> (See July 14th 07 for protocol). (both tubes are the same sample) with EcoR1 and Xba. <br><br />
1a- sample 1 - 5microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + 10microL ddH20<br><br />
1a- sample 2 - 15microL I0500 DNA + 1microL T4 ligase + 4microL 5X ligase buffer + NO WATER<br />
<br />
<br />
2- <b>Restrict Benny and Enny in Boo's </b>(in new unlabeled IGEM box-with other three boxes (DNA DNA and Buffers...) (with PstI/SpeI- there should be a protocol in master book within last week) <br><br />
<br />
*use 4microL "in boo" DNA of each sample <br />
<br />
<br />
3-Run Gel, Purify, Ligate I0500 into J61003 (that has already been restricted with EcoR1 and Xba), (freeze Benny and Enny gel purified SpeI/PstI) <BR><br />
4-Transform I0500 in J61003 into comp Hb101, sequence... rxn<br><br />
<br />
<br />
1.Extract the bands that have been cut out (are next to I0500 in box); Betty and Buddy from Aug 23rd AM gel,. <br><br />
<br />
Sequencing of D.B in B00 #2 and #7 has been done. I will update the results Friday evening. -ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ML, JG, NK <br><br />
Thanks justin-NK<br />
<br />
<b>LAB WORK </b><br />
<br />
Restrictions where done on I0500 and Enny in boo and Benny in Boo,<br />
No I0500 bands where found on the gel, Bands where seen for benny-boo and enny-boo However gel got mangled in the process of moving it from camera to lab so no gel purification was not done.<br />
<br />
S.T.D. (Stuff to do)<br />
The restriction on Enny in Boo and Benny in Boo needs to be done. Gel needs to be reran and gel purification needs to be done. So pretty much everything that we did tonight lol.<br />
<br />
Quote of the Day<br />
<br />
"I became president to lead not to read!!!" <br />
<br />
Attendance<br />
NK, JG, AF, ML <br />
PEACE!<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, NK, MC<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b> Can we meet at 10:30 instead of noon? -CZ<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JP,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,JG<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AL,JG,NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 31 ==<br />
<b>Schedule:</b><br />
<br />
CZ,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-08-25T00:13:32Z<p>CelineYZ: /* '''Become A Partner''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns in the global energy market there has been a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. In addition, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuels source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, cold starting, or volatile organic pollution standards. Blending butanol with gasoline also prevents modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''E.coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production in ''Clostridium acetobutylicum'' into ''E.coli''. Furthermore, we hope to increase the E.coli's tolerance to solvents such as butanol.<br />
<br />
<br />
Concurrently, we are looking into the use of a photoautotrophic bacterium, ''Chlorobium tepidum'', that we will also introduce butanol producing genes into. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as a terminal electron acceptors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees celcius (104 degrees fahrenheit for the yankees), and requires low light conditions for optimal growth. These bacteria grows well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
Figure 1 and 2 Images from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
More information on the technical details can be found '''[[Alberta/planb|here]]'''.<br />
<br />
== '''Become A Partner''' ==<br />
<br />
This summer, eleven members are participating in this very exciting project voluntarily without monetary stipend and fundraising to do the project from scratch. We would love to form a relationship with your organization's support to help achieve our goals in this exciting learning experience. <br><br />
<br />
If your organization is interested in becoming a partner and supporting our exciting learning adventures, contact our fundraising coordinator Michelle; mcchan[at]ualberta.ca or our mentor Dr. Michael Deyholos; deyholos[at]ualberta.ca<br><br />
<br />
'''Monetary'''<br />
<br />
<br />
[[Image: Alberta_UofAFofS.gif]][[Image: Alberta_AI.gif]][[Image: UofABiochem.jpg]]<br><br><br />
Alberta Research Council <br><br />
University of Alberta - Department of Biochemistry<br><br />
University of Alberta - Department of Pharmacology<br><br />
Trimline Design Centre Inc. <br><br />
ALberta Advanced Education & Technology <br><br />
<br />
<br />
<br />
'''Supplies'''<br />
<br />
<br />
[[Image: Alberta_AB.gif]]<br><br />
<br />
<br />
'''In-Kind'''<br />
<br />
<br />
[[Image: Alberta_biobasic.gif]]<br />
[[Image: Alberta_geneart.gif]]<br />
[[Image: Alberta_UofABS.gif]]<br><br />
<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor,''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta,''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta,''<br><br />
'''Dr. David Bressler''' - ''University of Alberta,''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta,''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta,''<br><br />
'''Dr. Donald Bryant''' - ''Penn State,''<br><br />
'''Dr. Gaozhong Shen''' - ''Penn State,''<br><br />
'''Amaya Garcia''' - ''Penn State,''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta,''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta,''<br><br />
'''Dr. Federick West'''-''University of Alberta,''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta,''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
'''Dr. Jeff Fuller''' - ''University of Alberta/Capital Health''<br><br />
'''Dr. Julia Foght''' - ''University of Alberta''<br><br />
<br />
<br />
<br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
== '''Discussion Board''' ==<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''Protocols''' ==<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
== '''Files''' ==<br />
<br />
Meeting minutes, agendas and action items can be found '''[[alberta/meeting files|here]]'''<br />
<br />
Other shared files can be found '''[[alberta/files|here]]'''. (If you have a file to post please contact Nick Glass)<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
<br />
== '''Edmonton''' ==<br />
<br />
For more on the city of Edmonton click [http://www.ualberta.ca/~mjl3/About.html here].<br />
<br />
For more info on the University of Alberta '''[http://www.ualberta.ca University of Alberta click here]'''<br />
<br />
== '''External Links''' ==<br />
<br />
'''Computational Modelling'''<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-08-17T03:59:27Z<p>CelineYZ: /* '''Become A Partner''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns in the global energy market there has been a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. In addition, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuels source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, cold starting, or volatile organic pollution standards. Blending butanol with gasoline also prevents modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''E.coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production in ''Clostridium acetobutylicum'' into ''E.coli''. Furthermore, we hope to increase the E.coli's tolerance to solvents such as butanol.<br />
<br />
<br />
Concurrently, we are looking into the use of a photoautotrophic bacterium, ''Chlorobium tepidum'', that we will also introduce butanol producing genes into. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as a terminal electron acceptors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees celcius (104 degrees fahrenheit for the yankees), and requires low light conditions for optimal growth. These bacteria grows well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
Figure 1 and 2 Images from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
More information on the technical details can be found '''[[Alberta/planb|here]]'''.<br />
<br />
== '''Become A Partner''' ==<br />
<br />
This summer, eleven members are participating in this very exciting project voluntarily without monetary stipend and fundraising to do the project from scratch. We would love to form a relationship with your organization's support to help achieve our goals in this exciting learning experience. <br><br />
<br />
If your organization is interested in becoming a partner and supporting our exciting learning adventures, contact our fundraising coordinator Michelle; mcchan[at]ualberta.ca or our mentor Dr. Michael Deyholos; deyholos[at]ualberta.ca<br><br />
<br />
'''Monetary'''<br />
<br />
<br />
[[Image: Alberta_UofAFofS.gif]][[Image: Alberta_AI.gif]][[Image: UofABiochem.jpg]]<br><br><br />
Alberta Research Council <br><br />
University of Alberta - Department of Biochemistry<br><br />
University of Alberta - Department of Pharmacology<br><br />
Trimline Design Centre Inc. <br><br />
ALberta Advanced Education & Technology <br><br />
<br />
<br />
<br />
'''Supplies'''<br />
<br />
<br />
[[Image: Alberta_AB.gif]]<br><br />
<br />
<br />
'''In-Kind'''<br />
<br />
<br />
[[Image: Alberta_biobasic.gif]]<br />
[[Image: Alberta_geneart.gif]]<br />
[[Image: Alberta_UofABS.gif]]<br><br />
<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor,''<br><br />
'''Dr. Jonathan Dennis''' - ''University of Alberta,''<br><br />
'''Dr. Perrin Beatty''' - ''University of Alberta,''<br><br />
'''Dr. David Bressler''' - ''University of Alberta,''<br><br />
'''Dr. Charles Lucy''' - ''University of Alberta,''<br><br />
'''Dr. Gregory Kiema''' - ''University of Alberta,''<br><br />
'''Dr. Donald Bryant''' - ''Penn State,''<br><br />
'''Dr. Gaozhong Shen''' - ''Penn State,''<br><br />
'''Amaya Garcia''' - ''Penn State,''<br><br />
'''Dr. Mark S. Peppler''' - ''University of Alberta,''<br><br />
'''Dr. James Harynuk''' - ''University of Alberta,''<br><br />
'''Dr. Federick West'''-''University of Alberta,''<br><br />
'''Dr. Rik Tykwinski'''-''University of Alberta,''<br><br />
'''Dr. Todd Lowary'''-''University of Alberta,''<br><br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br><br />
<br />
<br />
<br />
<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
== '''Discussion Board''' ==<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''Protocols''' ==<br />
<br />
[http://www.ualberta.ca/~mjl3/UofAIgemProtocols.pdf The Lab Protocols]<br />
<br />
== '''Files''' ==<br />
<br />
Meeting minutes, agendas and action items can be found '''[[alberta/meeting files|here]]'''<br />
<br />
Other shared files can be found '''[[alberta/files|here]]'''. (If you have a file to post please contact Nick Glass)<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
<br />
== '''Edmonton''' ==<br />
<br />
For more on the city of Edmonton click [http://www.ualberta.ca/~mjl3/About.html here].<br />
<br />
For more info on the University of Alberta '''[http://www.ualberta.ca University of Alberta click here]'''<br />
<br />
== '''External Links''' ==<br />
<br />
'''Computational Modelling'''<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-13T16:19:06Z<p>CelineYZ: /* August 13 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>Omniscient Notes:</b><br />
<br />
1- <b> Label Tubes Well </b> - There is nothing worse than having to redo your samples because you don't know what each tube is. Aside of our coding namings Benny, betty etc, <b>Label in detail</b>. Always <b>put the date</b> on the tubes so we can always refer to the master book if we're confused<br />
<br />
<b>MC available in AM to do some lab work - if needed please notify via email - MC</b><br />
<br />
I took out plates for I13453 this morning and there are big colonies to pick tonight.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AF, JG <br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK<br />
<br><br />
Hey guys i cant make it on 17,18 or 19. I have to go to seattle. Can someone switch with me please. Thanks -NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
== August 31 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-12T20:12:36Z<p>CelineYZ: /* August 12 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
1. got a hold of Meagan at MIT and she's sending a stub of IO500 bacteria. we should get it early next week I hope! <br><br />
<br />
<b> Lab Notes: </b><br />
<br />
1- organized all important samples into new box in -20 (all coding sequences in Boo) These will need to be restricted with Xba/PST for ligation into J61003 later <br><br />
2-started ligation on Diezel into Boo (samples at the front of the room on bench - need to transform tomorrow asap) <br><br />
3- autoclaved <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<b>Celine: wanna meet up at noon? AL</b><br />
<br />
<b> Lab Notes </b><br />
There was a misunderstanding about the Diezel Blade double digest. On Friday, we only did the digest but did not run a gel or perform gel extraction. So the ligation mixture from Saturday was discarded because B0034 was ligated to the double digest mixture directly.<br />
<br />
Today we ran the remaining double digest mixture; however it did not cut.<br />
<br />
Also we transformed Bba_I13453 (pBAD promoter by itself) just in case on Amp plates, which are in 37 degree incubator.<br />
<br />
<b> For tomorrow </b><br />
1. Take out I13453 plates and put in 4 degree fridge (Celine will do this early morning around 8am)<br />
2. Pick colonies for I13453 and grow in Amp-LB overnight<br />
3. Double-digest of Diezel, run gel, gel extraction, ligation with B0034.<br />
<br />
<b> Justin, can I swap with you for Monday because I may not make it on Thursday if we are doing labwork before the meeting? Email me to confirm. Thanks. - CZ </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br><br />
<br />
<b>MC available in AM to do some lab work - if needed please notify via email - MC</b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AF, JG <br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK<br />
<br><br />
Hey guys i cant make it on 17,18 or 19. I have to go to seattle. Can someone switch with me please. Thanks -NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
== August 31 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-11T03:29:12Z<p>CelineYZ: /* August 10 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
<b>Lab Notes</b><br />
<br />
Ran gel of pcr product no bands where observed. PCR reminants are stored in -20 freezer.<br />
<br />
Double digest of diesel blaze pst1 xba1, Stored in -20 freezer<br />
<br />
Gel already prepared on the second bench.<br />
<br />
Culture tubes have been wash but need to be autoclaved tommorrow<br />
<br />
ACTION ITEM: decide on promoter to transform we promoter list is on the mac computer. <br />
<br />
<b>Uplifting thought:</b><br />
<br />
A negative result is still a useful result. As long as it is within errors it is still valid.<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AF, JG <br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
== August 31 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-10T20:17:52Z<p>CelineYZ: /* August 16 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br><br />
2. PCR for <br><br />
a. Enny + B0034 miniprep 1 to 8 <br><br />
b. Betty colonies (10)<br><br />
c. Buddy colonies (10)<br><br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br><br />
<br />
For tomorrow<br><br />
1. run gel of PCR prodcut which are in the thermocycler � <br><br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br><br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br><br />
4. Make Amp plates (we only have 4 more)<br><br />
5. Autoclave test tubes <br><br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1830hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Made AMP plates<br><br />
Autoclaved test tubes<br><br />
Ran gel of ? <br><br />
Started O/N of Buddy & Betty<br><br />
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls<br><br />
<br />
James did a loving PCR of Benny in B0034 <br><br />
<br />
-MC, ED, JP, JB<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
<b> Meeting @ 1700hrs</b><br><br />
<br />
<b>Captain's Log, Stardate: 080307-1153</b> <br><br />
<br />
Minipreps of lifeforms Buddy and Betty <br><br />
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.<br><br />
-JB<br><br />
<br />
Made gel<br><br />
Digested (double) Buddy & Betty minipreps<br><br />
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer<br><br />
weekend to-do list<br><br />
-JP, ML, MC<br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<br />
<b>Lab Notes:</b><br />
<br />
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge<br><br />
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy<br><br />
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101<br><br />
<br />
-MC, JP, ML<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 5 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<b>Lab Notes:</b> <br><br />
<br />
1-transform I0500 into HB101 and XL10 gold <br><br />
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)<br><br />
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform... <br><br />
-MC, JP & ML (in spirit)<br />
<br />
<b> For Tomorrow </b><br />
<br />
1-gel purify Buddy and Betty <br><br />
2-ligate Buddy and Betty into Boo34 (overnite) <br><br />
3-start ONs of I0500 for mini on tuesday <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 6 ==<br />
<br />
<b> Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030</b><br />
<br />
<b>Lab Notes:</b> <br><br />
<br />
- JP, ML, AF, MC<br />
<br />
<b>Notes:</b><br />
<br />
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth <br><br />
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted <br><br />
3- began ligation reaction with B0034 (in back shaker at 13 degrees) <br><br />
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous <br><br />
<br />
<b>Fact of the Day</b><br />
<br />
A good ligation is like a hoola-hoop, they're both circular and only one is methylated <br><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 7 ==<br />
<br />
<b>Schedule:</b><br />
<br />
MC,VH,AF<br />
<br />
<b>Protocol for Today</b><br />
<br />
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube) <br><br />
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.<br> dilute (1)2microL into 8microL water and<br> (2)1microL into 9microL water. <br>Take the four samples and transform them (dilute in 600microL LB before puting into shaker.<br> Plate each transformant onto AMP plates in two dilutions: <br>(1)200microL on a plate<br> (2)50microL + 50microL LB. <br> <br />
<br />
<b>Lab Notes:</b><br />
<br />
No growth on Kan plates of I0500 in HB101 - therefore no O/N<br><br />
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells<br><br />
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20<br><br />
- labelling same for Buddy+BOO34<br />
<br />
- MC, VH, AF<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 8 ==<br />
<br />
Today is sweatpants day. Wear your favorite pair.<br />
<br />
<b>The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD </b><br />
<br />
<b>LAB NOTES:</b> <br><br />
<br />
1: transformed buAld_DH "Diezel Blaze" into XL10 gold need to grow up for mini tomorrow nite <br><br />
2: colony PCR on buddy and betty in Boo- reactions are in PCR machine (hopefully at 4 degreesC) <br><br />
3: started Chlorobium medium/concoction need some more compounds <br><br />
<br />
-JP,ED,AF<br />
<br />
<b>Schedule:</b><br />
<br />
ED,NK,JP<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 9 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED,AF,JG<br />
<br />
'''Lab Notes:'''<br />
-poured 1%-4% butanol-LB plates - added butanol to each individual plate as it was cooling. plates are on bench #3<br />
<br />
-ran gel of PCR of Buddy and Betty in Boo. Looks like all colonies tested were positive. Started overnights of positive colonies for glycerol stock and miniprep tomorrow.<br />
<br />
-set up PCR using VF and VR and 0.5 uL of 1/10 dilution of I0500 stock as template. Mg titration of 1,2,3 mM. Need to be run on gel tomorrow<br />
<br />
-started overnights of Diezel Blaze for miniprep and glycerol stocks<br />
<br />
-ED,MC,JG<br />
<br />
<br><br><br />
<b> Modeling meeting cancelled as Nick G. and Doug are both away - WM </b><br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 10 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, ML, JG <br><br />
<b>Thanks for taking my shift ML - MC<br><br />
Please note that if you cannot make a shift we have already discussed that you are to call the lab phone number which is in previous minutes - I can send it out with the next minutes just in case. - MC&JG</b><br />
<br />
'''Lab Notes:'''<br />
<br />
Minipreps and glycerol stocks of Buddy and Betty in Boo and Diezel Blaze were done and stored in box with green tape.<br />
<br />
Tonight: run I0500 PCRs on gel, digest Diezel Blaze to clone into Boo, start mutagenesis experiments, transform some new promoters<br />
<br />
This weekend: we can start cloning our genes into J61003 with this promoter and then swap the promoters later if we want (this was the reason for picking J61003), also we need to wash and autoclave culture tubes<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 11 ==<br />
<br />
<b>Schedule: 9:00am sharp as a needle</b> <br />
<br />
JP, AL, NK<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 12 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 13 ==<br />
<b>Schedule:</b><br />
<br />
ML,JG,JP<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 14 ==<br />
<b>Schedule:</b><br />
MC,JP,ML<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 15 ==<br />
<b>Schedule:</b><br />
CZ,AL,NK<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 16 ==<br />
<b>Schedule:</b><br />
<br />
JP,CZ,AL<br />
If lab can happen before meeting we can just leave for erins house after the meeting so you three should look into that<br />
JG<br />
<br />
I most likely cannot meet at the lab before the meeting, but if either Erin or Justin can. I would be willing to swap. Please let me know ASAP, thanks.-CZ<br />
<br />
Post-meeting video/board game party at Erin's house if there is interest.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 17 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AF, JG <br><br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
ED, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 19 ==<br />
<b>Schedule:</b><br />
<br />
CZ, AL, NK<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 20 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 21 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 22 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 23 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 24 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 25 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 26 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 27 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 28 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 29 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 30 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
== August 31 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/AlbertaAlberta2007-08-05T00:36:05Z<p>CelineYZ: /* '''Become A Partner''' */</p>
<hr />
<div><center>[[Image: headbanner.jpg]] <br />
<br><br />
''The Offical Wiki of the 2007 University of Alberta iGEM Team'' <br><br />
''Edmonton, Alberta, Canada </center>''<br />
<br />
== '''Background: Biofuels'''==<br />
<br />
<br />
With growing concerns in the global energy market there has been a continual push for the development of renewable energy sources. Specifically, two fuels have dominated the media: biodiesel and ethanol. However, both of these fuels have shortcomings in terms of being a viable fuel source.<br />
<br />
<br />
Biodiesel is a fuel produced from the vegetable oils of crops that can be used in an engine system very similar to a traditional diesel engine. However, vegetable oils in crops only make up a small portion of the biomass of the plants, ultimately producing low yields of fuel per acre of crops. As such, it is more economically sound to use these resources for food production.<br />
<br />
<br />
There has been huge attention to the use of ethanol in the typical auto cycle engine. In many places it is already being blended with gasoline to create a hybrid fuel source. However, running an engine on pure ethanol is beset by several major obstacles. Firstly, ethanol is miscible with water at any concentration, which creates long term storage corrosion issues. In addition, ethanol engines must be designed to expect water vapor unlike their gasoline counterparts. Ethanol also has significantly different thermodynamic properties than gasoline such as a lower energy density and different vapor properties, which would reduce the economical advantage of using ethanol as a primary fuels source.<br />
<br />
<br />
We propose using a different fuels source to eventually replace gasoline, butanol. Butanol is a superior to ethanol as a replacement for petroleum gasoline. With a low vapor pressure, high energy density, and a gasoline-like octane rating, it can be blended into existing gasoline at much higher proportions than ethanol without compromising performance, mileage, cold starting, or volatile organic pollution standards. Blending butanol with gasoline also prevents modifications of the fuel-air ratio and modifications to the fuel system. Butanol also is immiscible with water at concentrations higher than 7%, alleviating storage concerns.<br />
<br />
<br />
More information on the summary of biofuels viability and the inspiration to our project can be found '''[[Alberta/background|here]]'''.<br />
<br />
== '''The Project: Plan B''' ==<br />
<br />
We propose the use of butanol as the leading biofuel for use in internal combustion engines. Specifically, we intend to genetically engineer ''E.coli'' bacteria to convert biomasses into butanol for use as an energy source. This will be accomplished by introducing the genes responsible for butanol production in ''Clostridium acetobutylicum'' into ''E.coli''. Furthermore, we hope to increase the E.coli's tolerance to solvents such as butanol.<br />
<br />
<br />
Concurrently, we are looking into the use of a photoautotrophic bacterium, ''Chlorobium tepidum'', that we will also introduce butanol producing genes into. ''Chlorobium tepidum'' is a green sulfur bacterium that is strictly anaerobic and uses sulfur compounds as a terminal electron acceptors. ''Chlorobium tepidum'' is a moderately thermophillic bacterium, growing at 40 degrees celcius (104 degrees fahrenheit for the yankees), and requires low light conditions for optimal growth. These bacteria grows well in a defined medium, utilizing the reverse/reductive tricarboxylic acid (TCA) cycle to build up carbohydrates from carbon dioxide. For a more comprehensive overview of ''Chlorobium tepidum'' go to http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
[[Image:spacer.jpeg|left]][[Image:Alberta_micrograph.gif|thumb|300px|left|Figure 1:''Chlorobium tepidum'' Transmission Micrograph Image]]<br />
[[Image:Alberta_gramstain.jpg|thumb|330px|left|Figure 2:''Chlorobium tepidum'' Standard Light Microscope Image]]<br />
<br />
<br><br><br><br><br><br><br><br><br><br><br />
Figure 1 and 2 Images from http://www.bmb.psu.edu/faculty/bryant/lab/chlorobiumtepidum/index.html<br />
<br />
The advantage of manipulating this organism for butanol production is a matter of energy input. Rather than having to utilize vast amounts of food stocks, such as grains or sugars, a photoautotroph will fix carbon dioxide into the complex carbohydrates required for butanol production. Theoretically, if the only fuel on our planet was butanol, and it was produced in this manner, there would be little net carbon dioxide produced.<br />
<br />
More information on the technical details can be found '''[[Alberta/planb|here]]'''.<br />
<br />
== '''Become A Partner''' ==<br />
<br />
'''Monetary'''<br />
<br />
<br />
[[Image: Alberta_UofAFofS.gif]]<br />
[[Image: Alberta_AI.gif]]<br><br><br />
University of Alberta Bookstore<br> <br />
Alberta Research Council <br><br />
University of Alberta - Department of Biochemistry<br><br />
University of Alberta - Department of Pharmacology<br><br />
<br />
'''Supplies'''<br />
<br />
<br />
[[Image: Alberta_AB.gif]]<br><br />
<br />
<br />
'''In-Kind'''<br />
<br />
<br />
[[Image: Alberta_biobasic.gif]]<br />
[[Image: Alberta_geneart.gif]]<br />
[[Image: Alberta_UofABS.gif]]<br><br />
<br />
<br />
'''Advice'''<br />
<br />
<br />
'''Andrew Hessel''' - ''Alberta Ingenuity Mentor,''<br />
'''Dr. Jonathan Dennis''' - ''University of Alberta,''<br />
'''Dr. Perrin Beatty''' - ''University of Alberta,''<br />
'''Dr. David Bressler''' - ''University of Alberta,''<br />
'''Dr. Charles Lucy''' - ''University of Alberta,''<br />
'''Dr. Gregory Kiema''' - ''University of Alberta,''<br />
'''Dr. Donald Bryant''' - ''Penn State,''<br />
'''Dr. Gaozhong Shen''' - ''Penn State,''<br />
'''Amaya Garcia''' - ''Penn State,''<br />
'''Dr. Mark S. Peppler''' - ''University of Alberta,''<br />
'''Dr. James Harynuk''' - ''University of Alberta,''<br />
'''Desiree Schell''' - ''University of Alberta CJSR''<br />
<br />
<br />
<br />
If your organization is interested in becoming a partner and supporting our exciting learning adventures, contact our fundraising coordinator Michelle; mcchan[at]ualberta.ca or our mentor Dr. Michael Deyholos; deyholos[at]ualberta.ca<br />
<br />
A list of experimental supplies we need can be found '''[[supplies|here]]'''<br />
<br />
== '''Lab Book and Calendar''' ==<br />
[[image: Alberta_butapipettips.jpg]][[image: Alberta_arrow.jpg]] [[image: Alberta_Labbook.jpg]] <br><br />
<br />
Butanerd Event Calendar can be found below:<br />
<br />
[[Alberta/Calender/July|July 2007]]<br />
<br />
[[Alberta/Calender/August|August 2007]]<br />
<br />
[[Alberta/Calender/September|September 2007]]<br />
<br />
== '''Discussion Board''' ==<br />
<br />
'''[http://butanerds.myfreeforum.org The Butanerd Online Forum]''' is up and running. Feel free to post and check for posts here. Make sure you register!<br />
<br />
== '''Protocols''' ==<br />
<br />
[http://www.ualberta.ca/~nglass/IGEM_Web/UofA%20iGEM%20Protocols.pdf The Lab Protocols]<br />
<br />
== '''Files''' ==<br />
<br />
Meeting minutes, agendas and action items can be found '''[[alberta/meeting files|here]]'''<br />
<br />
Other shared files can be found '''[[alberta/files|here]]'''. (If you have a file to post please contact Nick Glass)<br />
<br />
== '''The Team''' ==<br />
<br />
<center>[[Image: Alberta_team.jpg]] </center><br />
<br><br />
<br />
Our team has a rich background in biology, biochemistry and engineering. To compliment our diversity we also have advisors who have a wealth of knowledge in research and applications of genetic engineering. For more information about the group, check out the '''[[Alberta/Members|University of Alberta's iGEM Team Members Page]]'''.<br />
<br />
Our University of Alberta Student Group Constitution can be found '''[[alberta/constitution|here]]'''<br />
<br />
<br />
== '''Edmonton''' ==<br />
<br />
For more on the city of Edmonton click [http://www.ualberta.ca/~mjl3/About.html here].<br />
<br />
For more info on the University of Alberta '''[http://www.ualberta.ca University of Alberta click here]'''<br />
<br />
== '''External Links''' ==<br />
<br />
'''Computational Modelling'''<br />
<br />
[http://www.systems-biology.org/cd Cell Designer Homepage]<br />
<br />
[http://www.biomodels.org Biomodels Home Page]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/AugustAlberta/Calender/August2007-08-02T03:31:04Z<p>CelineYZ: /* August 1 */</p>
<hr />
<div>=='''August'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/August|August 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td>[[Alberta/Calender/August#August_1|1]]</td><br />
<td>[[Alberta/Calender/August#August_2|2]]</td><br />
<td>[[Alberta/Calender/August#August_3|3]]</td><br />
<td>[[Alberta/Calender/August#August_4|4]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_5|5]]</td><br />
<td>[[Alberta/Calender/August#August_6|6]]</td><br />
<td>[[Alberta/Calender/August#August_7|7]]</td><br />
<td>[[Alberta/Calender/August#August_8|8]]</td><br />
<td>[[Alberta/Calender/August#August_9|9]]</td><br />
<td>[[Alberta/Calender/August#August_10|10]]</td><br />
<td>[[Alberta/Calender/August#August_11|11]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_12|12]]</td><br />
<td>[[Alberta/Calender/August#August_13|13]]</td><br />
<td>[[Alberta/Calender/August#August_14|14]]</td><br />
<td>[[Alberta/Calender/August#August_15|15]]</td><br />
<td>[[Alberta/Calender/August#August_16|16]]</td><br />
<td>[[Alberta/Calender/August#August_17|17]]</td><br />
<td>[[Alberta/Calender/August#August_18|18]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_19|19]]</td><br />
<td>[[Alberta/Calender/August#August_20|20]]</td><br />
<td>[[Alberta/Calender/August#August_21|21]]</td><br />
<td>[[Alberta/Calender/August#August_22|22]]</td><br />
<td>[[Alberta/Calender/August#August_23|23]]</td><br />
<td>[[Alberta/Calender/August#August_24|24]]</td><br />
<td>[[Alberta/Calender/August#August_25|25]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/August#August_26|26]]</td><br />
<td>[[Alberta/Calender/August#August_27|27]]</td><br />
<td>[[Alberta/Calender/August#August_28|28]]</td><br />
<td>[[Alberta/Calender/August#August_29|29]]</td><br />
<td>[[Alberta/Calender/August#August_30|30]]</td><br />
<td>[[Alberta/Calender/August#August_31|31]]</td><br />
<td></td><br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
To [[Alberta/Calender/July|July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== August 1 ==<br />
<br />
<b>Schedule:</b><br />
CZ,NG,VH<br />
<br />
<b>General Notes:</b><br />
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.<br />
2. PCR for <br />
a. Enny + B0034 miniprep 1 to 8<br />
b. Betty colonies (10)<br />
c. Buddy colonies (10)<br />
Follow James' protocol. Also see blackboard for James' comment about PCR.<br />
<br />
For tomorrow<br />
1. run gel of PCR prodcut which are in the thermocycler �<br />
2. check restreaked single colonies of the Betty and Buddy colony PCR<br />
3. grow up Betty and Buddy overnight and for miniprep and ligation later<br />
4. Make Amp plates (we only have 4 more)<br />
5. Autoclave test tubes<br />
<br />
Quotes of the day:<br />
We are tired.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 2 ==<br />
<br />
Modeling Meeting @ 1800hrs in CAB373<br><br />
General Meeting @ 1900hrs in CAB373<br><br />
Party afterward's @ Garneau pub<br><br />
<br />
<b>Schedule:</b><br />
<br />
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like.<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 3 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough<br><br />
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads. <br />
<br />
<br />
[[Alberta/Calender/August#August|to the top]]<br />
<br />
== August 4 ==<br />
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== August 5 ==<br />
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== August 7 ==<br />
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== August 14 ==<br />
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== August 15 ==<br />
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== August 26 ==<br />
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== August 27 ==<br />
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== August 28 ==<br />
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== August 29 ==<br />
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== August 30 ==<br />
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== August 31 ==<br />
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[[Alberta/Calender/August#August|to the top]]<br />
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[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/July|To July 2007]]<br><br />
To [[Alberta/Calender/September|September 2007]]<br></div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/JulyAlberta/Calender/July2007-07-29T22:42:34Z<p>CelineYZ: /* July 29 */</p>
<hr />
<div><b>Schedule Legend:</B><br />
JG: Jason, <br />
Mc: Michelle,<br />
ED: Erin,<br />
NG: Nick G.,<br />
NK: Nik K.,<br />
CZ: Celine,<br />
AF: Adam,<br />
Al: Alex,<br />
JB: Jori,<br />
JP: Justin,<br />
VH: Veronica,<br />
<br />
<br />
=='''July'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<br />
<br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/July|July 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_1|1]]</td><br />
<td>[[Alberta/Calender/July#July_2|2]]</td><br />
<td>[[Alberta/Calender/July#July_3|3]]</td><br />
<td>[[Alberta/Calender/July#July_4|4]]</td><br />
<td>[[Alberta/Calender/July#July_5|5]]</td><br />
<td>[[Alberta/Calender/July#July_6|6]]</td><br />
<td>[[Alberta/Calender/July#July_7|7]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_8|8]]</td><br />
<td>[[Alberta/Calender/July#July_9|9]]</td><br />
<td>[[Alberta/Calender/July#July_10|10]]</td><br />
<td>[[Alberta/Calender/July#July_11|11]]</td><br />
<td>[[Alberta/Calender/July#July_12|12]]</td><br />
<td>[[Alberta/Calender/July#July_13|13]]</td><br />
<td>[[Alberta/Calender/July#July_14|14]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_15|15]]</td><br />
<td>[[Alberta/Calender/July#July_16|16]]</td><br />
<td>[[Alberta/Calender/July#July_17|17]]</td><br />
<td>[[Alberta/Calender/July#July_18|18]]</td><br />
<td>[[Alberta/Calender/July#July_19|19]]</td><br />
<td>[[Alberta/Calender/July#July_20|20]]</td><br />
<td>[[Alberta/Calender/July#July_21|21]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_22|22]]</td><br />
<td>[[Alberta/Calender/July#July_23|23]]</td><br />
<td>[[Alberta/Calender/July#July_24|24]]</td><br />
<td>[[Alberta/Calender/July#July_25|25]]</td><br />
<td>[[Alberta/Calender/July#July_26|26]]</td><br />
<td>[[Alberta/Calender/July#July_27|27]]</td><br />
<td>[[Alberta/Calender/July#July_28|28]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_29|29]]</td><br />
<td>[[Alberta/Calender/July#July_30|30]]</td><br />
<td>[[Alberta/Calender/July#July_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<br />
<br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== July 1 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 2 ==<br />
<br />
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[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 3 ==<br />
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[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 4 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.<br />
<br />
-ED, JG, ML, MC, VH, AL, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 6 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformation of E1010 did not work, it is in a Kan vector. SMRT.<br />
<br />
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 7 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.<br />
<br />
-JP,JB,AL<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 8 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.<br />
<br />
Pour Kan and Amp plates. Make TBE and TAE buffer.<br />
<br />
-ED, JP, MC, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 9 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.<br />
<br />
-ED, VH, JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 10 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.<br />
<br />
-ED, MC, AF, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 11 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.<br />
<br />
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.<br />
<br />
<center> [[image: Alberta_e1010xlgold_july11.jpg]] <br />
<br />
E1010 XLGOLD 10 July 11</center><br />
<br />
-ED, MC, NG, JG, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 12 ==<br />
<br />
'''General Notes:'''<br />
<br />
Calender up and running.<br />
<br />
Meeting tonight at 7:00 in CSC 2-49.<br />
<br />
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!!<br />
-Justin<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.<br />
<br />
Streak for single colonies of E1010, A0500, J61003.<br />
<br />
Purification of gel slice of digested J61003.<br />
<br />
<br />
<center> [[image: Alberta_e1010DH5a_july12.jpg]] <br />
<br />
E1010 DH5a July 12</center><br />
<br />
-ED, JG, AL, JB, MC, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 13 ==<br />
<br />
'''General Notes:'''<br />
<br />
Everyone who is coming tonight please be present 7:00pm sharp<br />
<br />
'''Lab Notes:'''<br />
<br />
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.<br />
<br />
-ED, JB<br />
<br />
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"<br><br />
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)<br><br />
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL<br><br />
Made some Kan plates<br><br />
<br />
-MC, JP, JG, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 14 ==<br />
<b>Schedule:</b><br />
<br />
AL,JP,NK<br />
<br />
<b>General Notes:</b><br />
<br />
If you are scheduled during the day please be in the lab at 1230hrs (PM) <br />
Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911<br />
<br />
Things to be continued today (CZ)<br><br />
1. Put LB plates in 4 degree fridge <br><br />
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control<br><br />
3. Xba1/EcoRI double digest <br><br />
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour. <br><br />
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.<br><br />
<br />
If you need anything else call me(Celine). Number on the blackboard. Thank you.�<br />
<br />
<b/>Lab Notes:</b><br />
JP, AL, NK, JB(?) <br><br />
<br />
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20 <br><br />
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20 <br><br />
3. Placed Kan plates at +4 <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 15 ==<br />
<b>Schedule:</b><br />
<br />
Jp,JB,AL<br />
<br />
<b>General Notes:</b><br />
<br />
Options: <br><br />
<br />
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)<br><br />
2. gel purify J61003 restriction products (promoter restriction sample) <br><br />
3. fundraising letters! <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow<br><br />
2. glycerol stocks of Chlorobium tepidum<br><br />
3. finished grant application forms<br><br />
- JP, AL, NG, MC<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 16 ==<br />
<b>Schedule:</b><br />
<br />
CZ,VH,AF<br />
<br />
<b>General Notes:</b><br />
<br />
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP) <br><br />
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining <br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed gel extraction of E,X digested J61003. (in box in -20)<br />
<br />
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.<br />
<br />
-VH, ED, CZ<br />
<br />
<br />
'''Quotes of the Day:'''<br />
<br />
"If I had a remote, I'd change the cd."<br />
<br />
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it<br />
<br />
"I would never drive this tired. But I WILL operate chemicals."<br />
<br />
Anonymous<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 17 ==<br />
<b>Schedule:</b><br />
<br />
ED,VH,AF<br />
<br />
'''Lab Notes:'''<br />
<br />
Gel extracted E1010 from yesterday's slices.<br />
<br />
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post. <br><br />
<b> It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC </b> <br><br />
<br />
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED<br />
<br />
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.<br />
<br />
-ED,JG,VH,AF<br />
<br />
'''Quote of the Day'''<br />
<br />
"Cancer is an urban myth."<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,MC,NG<br />
<br />
<b>Lab Notes:</b><br />
<br />
Transformed Ligations 1,2,3,4 into XL10 Gold Competent cells <br><br />
Plated 1ul,10 ul, and 200ul on AMP <br><br />
Incubated @ 37celsius overnight<br><br />
<br />
-MC, NG, CZ<br />
<br />
<b>Quote(s) of the Day:</b><br />
<br />
"Asuuuuuuuuuuuum!!" <br><br />
In response to the awesome; "You're wayyyy to exciting for me right now" <br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 19 ==<br />
<br />
Meeting at 7:00, CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
ED,MC,JG<br />
<br />
<b>General Notes</b><br />
<br />
New Schedule is up<br><br />
check out the jar o' love @ the front :) <br><br />
<br />
<b>Lab notes</b><br />
<br />
Started 8X5mL overnights to test colonies of the RFP ligation. Hopefully JB will be available to do minipreps tomorrow. Miniprep protocol is tucked in Master Book.<br />
<br />
-ED,JG,MC<br />
<br />
<b>Quote of the Day </b><br />
<br />
"There is a pee throw up smell it the washroom"<br />
<br />
Reply<br />
<br />
"Someone must be pregnant"<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 20 ==<br />
<b>Schedule:</b><br />
<br />
JG,ED,MC <br><br />
<br />
<b>General Notes:</b><br><br />
<br />
There is now a Jar O' Love in the lab; Please do not drink chemicals from said jar o'love. Check it out - it's at the front on the instructors bench. It's to give shout outs for people who are rocking out and 'taking one for the team'. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprep Lig3 (8 samples) overnights - JB<br><br />
Made 1L LB broth (16 bottles) at the back to be put away in the sterile cupboards tomorrow <br><br />
Ran Gel of JB's Minipreps after digestion with Xba and Spe. All of the colonies were positive (band at 1.2 kB).<br><br />
Did some dishes - we should learn how to use the dishwasher<br><br />
<br />
- MC, ED, JG <br><br />
<br />
<b>Quote(s) of the day:</b><br><br />
<br />
"Always use protection when using the Jar O' Love"<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 21 ==<br />
<b>Schedule:</b><br />
<br />
JP,NK,AL<br />
<br />
<b>General Notes:</b><br />
<br />
<br />
Since it doesn't look like Nik will be showing up, maybe just call Alex and see if it works for him.<br />
<br />
All of the ligations of RFP into J61003 turned out. They are labelled JB July 19 1-8 in the DNA box. Transform 1 of them into XL10 Gold and plate on Amp with Tetracycline plates. You will have to add the Tet to the plates beforehand. I don't know what concentraion, but James suggested 1 ug/mL. Try plating different amounts.<br />
<br />
The reason that I am making these suggestions is that I read that XL10Golds are Tet resistant, so we can maybe induce the expression of RFP with the Tet because it is under the Tet promotor and then we can see the RFP.<br />
<br />
As always, call me if you need anything.<br />
<br />
-ED<br />
<br />
<b>Lab Notes:</b><br />
<br />
1. got into building <br><br />
2. waited for peers - no shows <br><br />
3. tried to break into room by climbing through roof (cement goes all the way up) <br><br />
4. went home... will complete work tomorrow - sorry i'll get swipe access asap... <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 22 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,AL,NG<br />
<br />
<b>General Notes:</b><br />
<br />
How does 7:00pm sound?<br />
<br />
<b>Laboratory Notes:</b><br />
<br />
Transformed XL10 gold cells with J61003 (with RFP ligated in). Split cell aliquot into two 50microL aliquots and added 1microL DNA from ligation sample 1 and from ligation sample 4. Covered AMP plates with 50microL 1microg/ml tet, spread, let sit. Plated (1) 50microL sample and (1) 5microL + 45microL LB onto Tet/Amp plates for #'s 1&4. They are in 37C incubator. The remaining transformed cells are in +4C labeled July 22. <br><br />
<br />
<b>Serious Quote of the Month</b><br />
<br />
<b>Faust</b><br><br />
Then shall I see, with vision clear, <br><br />
How secret elements cohere, <br><br />
And what the universe engirds, <br><br />
And give up huckstering with words <br><br />
<br />
-Johann Wolfgang von Goethe <br><br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,CZ,JG<br />
<br />
<b>Laboratory Notes:</b><br><br />
1. Add more Tet on ligation plates from July 22. Hopefully red colonies will appear!<br><br />
2. Transformation of XL10Gold with<br><br />
- Butanol CoA dehydrogenase (1 in 1000 dilution)<br><br />
- Butenoyl CoA dehydrogenase (1 in 1000 dilution)<br><br />
- J23018 (RFP positive control)<br><br />
<br />
20 and 100 microlitres of transformed culture plated on Amp plate and incubate at 37 degrees<br><br />
<br />
The leftover mixture is stored in 4 degree fridge in case of no growth.<br><br />
3. Repeate overnight ligation from July 22. The exact amounts of insert, vector etc. are on the blackboard.<br><br />
<br />
For tomorrow:<br><br />
1. Pick colonies for transformed plates.<br><br />
2. Plate ligation mixtures with appropriate amount of Tet.<br><br />
3. Observe red colonies from the July 22 ligation plates if the cells are still alive.<br><br />
<br />
Memorable Quote:<br><br />
Don't force it. Let it come out naturally.<br><br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,NG<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed I0500 (arabinose promotor) into HB101 competent cells. Plated 2, 20, 200 uL on Kan.<br />
<br />
Transformed ligations from yesterday into XL10 gold. Plated 100 uL on Amp.<br />
<br />
RFP control from yesterday worked, but sadly our ligations are not expressing any RFP. It was still exciting to see red control cells. It seems as if the Tet promotor is induced at a low level without any Tet.<br />
<br />
Started 4X5ml overnights of enoyl coA hydratase and butyryl coa DH. There seems to be some confusion with the naming of these proteins. These are the names that I have used in the BioBricks. We will be able to tell if they are correctly labelled when we do the digests. Enoyl coa hydratase is about 840 bp and butyryl coa DH is about 1200 bp.<br />
<br />
Note on agarose gels: Use only the wide combs (8 lanes) because we are having to many problems with the narrow lanes. This may mean that you will need to pour multiple gels (oh my). Also, try and run two ladders per gel until we get the ladder problem sorted out. Also, make sure that you zoom in on the gel when taking a picture.<br />
<br />
Note on Tet: should be stored 12.5 mg/mL in 50% ethanol, 50% water solution. This is a 1000X solution. Keep this in mind when using Tet again.<br />
<br />
-ED, AF, James<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Jp,MC,VH<br />
<br />
<b>General Notes:</b><br />
<br />
Use of wide combs- 5microL of sample works well with 3.5microL of ladder. Make sure you stir up the ladder before use!<br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed minipreps of enoylcoa hydratase and butyryl coa dh. They are in the new box with the green tape. I think that there are two types of digests we should be doing: one to make sure we have the right stuff (digest with EcoR1 and Pst1) and one to clone in the RBS. To clone in the RBS, cut B0034 with Eco and Spe and cut the enoyl coa hydratase and butyryl coa dh with Eco and Xba.<br />
<br />
Check to see if any colonies have grown on I0500 plates in incubator. There weren't any visible on Adam's transformations this morning. James also transformed some this morning, so they may be hard to see. If there are colonies, start overnights and see if Jori can do minipreps tomorrow. If Jori can't make it, get ahold of me and I can. AND if there are no colonies, just post here and I'll go check the plates in the morning.<br />
<br />
I also forgot to start glycerol stocks of our enoly coa hydratase and butyryl coa dh. If you guys could restart some overnights from the plates in the fridge so I can do it tomorrow that would be great.<br />
<br />
I had to rush to work this morning and didn't get any of this written in the lab book. So you guys can make fun of me or whatever. I guess I belong in the Science Hall of Shame.<br />
<br />
-ED <br><br />
<br />
Enny = Enoly CoA Hydratase<br><br />
Benny = Butytyl CoA DH<br />
<br />
Checked plates<br><br />
- Adam - no growth: parafilmed and put in 4 fridge<br><br />
- James'? J23018: red colonies parafilmed and put in 4 fridge (was this the control?)<br><br />
<br />
Overnights of Benny & Enny<br><br />
- 3x 5mlLB each <br><br />
<br />
Restrictions<br><br />
- B0034 with SPE & ECO<br><br />
- Enny with Xba & ECO<br><br />
-Benny with Xba & ECO<br><br />
These are stored in the -20 freezer<br><br />
<br />
- MC, NK, VH, NG, JP<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- Glycerol stocks of Enny & Benny (these colonies are likely different from the transformations as it was not marked out which colonies were picked for transformations)<br><br />
- Gels for restrictions of B0034, Enny & Benny<br />
- cut out the bands; gel extractions? (if there is time)<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 26 ==<br />
<br />
Meeting at 7:00, Room CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
AL,VH,ED<br />
<br />
<b>Lab Book</b><br />
<br />
Ran gel of "Enny", "Benny" and Boo IV against 1 kb ladder.<br />
<br />
Cleared up glasswares in the autoclave.<br />
<br />
Parafilm all unparafilmed plates.<br />
<br />
Checked out red fluorescence of J23018.<br />
<br />
Attempted working on fundraising letters (?)<br />
<br />
-ED, VH, AL, JG, MC, AF, Doug<br />
<br />
<b>Favourite "F" word of the day:</b><br />
<br />
fulcrum<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,MC,NK<br><br />
The theme is Foul music Friday - bring any bad music you have (or embarassed that you have in your collection)<br />
<br />
<b>General Notes:</b><br />
<br />
1. This is whats on the agenda for this evening:<br />
[[Image: Alberta_promoterprob.jpg]]<br />
<br />
<b>Lab book:</b><br />
<br />
Digest Enny(1,4), Benny (1,2) B0034(2,4) with PST, XBA, SPE<br><br />
Enny with PST and XBA<br><br />
Benny with PST and XBA<br><br />
B0034 with PST and SPE<br><br />
O/N of Benny and Enny glycerol stock (3 each)<br><br />
<br />
'''''Note the 4 degree fridge at the back is out of commision. All iGEM materials have been moved to the room where we take pictures of gels. The door labelled IGEM.''''' Nik is here on saturday and Celine can show on sunday.<br><br />
<br />
- JG, NK, MC, JP, and James (cos he loves us) <br />
<br />
<br />
<b>Jame's Fun Tip of the Day:</b><br />
If you want to run a gel for a shorter period of time ad the EtBR to the gel rather than the buffer. You can add ~4ul to the gel rather than 10ul to the buffer. <br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JB,NK<br />
<br />
<b>Meeting @ 1300hrs - MC on behalf of CZ</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Glycerol stocks of Enny & Benny from the O/N<br><br />
- Gels of Enny, Benny & B0034 (put the EtBr in the gel)<br><br />
- Ligations of Enny+Benny into B0034 with T4 ligase; leave O/N in incubator at the back room (near the phone) set at 13degrees celsius<br />
<br />
<b>Lab Book</b><br />
<br />
Finished making glycerol stocks of Enny and Benny from the O/N. Followed Erin's protocols except the Flash Freeze step.<br><br />
<br />
Ran agarose gel of Benny, Enny, b0034 digest.<br> <br />
No DnA ladder (NEVER DO IT AGAIN). <br><br />
Ran at 90V for 20 Min, run longer for next time.<br><br />
<br />
Cut Benny band (1200BP) and B0034, need to redigest Enny tomorrow<br><br />
Gel extration using QIAGEN kit<br><br />
<br />
Ligation , control only with ligase, <br><br />
Stored at 13 degrees celsius overnight <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,AF,NG<br><br />
<br />
<b>As agreed on Thursday meeting at noon or 12</b><br />
<br />
<b>General Notes:</b><br />
<br />
1. Transformation of ligation (Benny and B0034( into XL10 Gold <br> on Amp plates; incubate at 37 degrees. Hopefully we have colonies to pick tomorrow.<br />
2. Double digest of Enny (Pst/Xba) and B0034 (Pst/Spe) repeated; Enny band (800bp) and B0034 (2.7kb) bands cut out and purified; overnight ligation mixture at 13 degree incubator at the back of the room<br />
<br />
<br />
For tomorrow:<br />
1. Pick colonies of Benny and B0034<br />
2. Transformation of Enny and B0034<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,JP,JG,<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 31 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,CZ,MC<br />
<br />
HORRAY Celine can take over for you Erin<br />
-JG<br />
<br />
Thanks Celine!<br />
<br />
-ED<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/June|To June 2007]]<br><br />
[[Alberta/Calender/August|To August 2007]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/JulyAlberta/Calender/July2007-07-29T02:42:37Z<p>CelineYZ: /* July 29 */</p>
<hr />
<div><b>Schedule Legend:</B><br />
JG: Jason, <br />
Mc: Michelle,<br />
ED: Erin,<br />
NG: Nick G.,<br />
NK: Nik K.,<br />
CZ: Celine,<br />
AF: Adam,<br />
Al: Alex,<br />
JB: Jori,<br />
JP: Justin,<br />
VH: Veronica,<br />
<br />
<br />
=='''July'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<br />
<br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/July|July 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_1|1]]</td><br />
<td>[[Alberta/Calender/July#July_2|2]]</td><br />
<td>[[Alberta/Calender/July#July_3|3]]</td><br />
<td>[[Alberta/Calender/July#July_4|4]]</td><br />
<td>[[Alberta/Calender/July#July_5|5]]</td><br />
<td>[[Alberta/Calender/July#July_6|6]]</td><br />
<td>[[Alberta/Calender/July#July_7|7]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_8|8]]</td><br />
<td>[[Alberta/Calender/July#July_9|9]]</td><br />
<td>[[Alberta/Calender/July#July_10|10]]</td><br />
<td>[[Alberta/Calender/July#July_11|11]]</td><br />
<td>[[Alberta/Calender/July#July_12|12]]</td><br />
<td>[[Alberta/Calender/July#July_13|13]]</td><br />
<td>[[Alberta/Calender/July#July_14|14]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_15|15]]</td><br />
<td>[[Alberta/Calender/July#July_16|16]]</td><br />
<td>[[Alberta/Calender/July#July_17|17]]</td><br />
<td>[[Alberta/Calender/July#July_18|18]]</td><br />
<td>[[Alberta/Calender/July#July_19|19]]</td><br />
<td>[[Alberta/Calender/July#July_20|20]]</td><br />
<td>[[Alberta/Calender/July#July_21|21]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_22|22]]</td><br />
<td>[[Alberta/Calender/July#July_23|23]]</td><br />
<td>[[Alberta/Calender/July#July_24|24]]</td><br />
<td>[[Alberta/Calender/July#July_25|25]]</td><br />
<td>[[Alberta/Calender/July#July_26|26]]</td><br />
<td>[[Alberta/Calender/July#July_27|27]]</td><br />
<td>[[Alberta/Calender/July#July_28|28]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_29|29]]</td><br />
<td>[[Alberta/Calender/July#July_30|30]]</td><br />
<td>[[Alberta/Calender/July#July_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<br />
<br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== July 1 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 3 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 4 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.<br />
<br />
-ED, JG, ML, MC, VH, AL, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 6 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformation of E1010 did not work, it is in a Kan vector. SMRT.<br />
<br />
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 7 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.<br />
<br />
-JP,JB,AL<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 8 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.<br />
<br />
Pour Kan and Amp plates. Make TBE and TAE buffer.<br />
<br />
-ED, JP, MC, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 9 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.<br />
<br />
-ED, VH, JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 10 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.<br />
<br />
-ED, MC, AF, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 11 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.<br />
<br />
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.<br />
<br />
<center> [[image: Alberta_e1010xlgold_july11.jpg]] <br />
<br />
E1010 XLGOLD 10 July 11</center><br />
<br />
-ED, MC, NG, JG, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 12 ==<br />
<br />
'''General Notes:'''<br />
<br />
Calender up and running.<br />
<br />
Meeting tonight at 7:00 in CSC 2-49.<br />
<br />
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!!<br />
-Justin<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.<br />
<br />
Streak for single colonies of E1010, A0500, J61003.<br />
<br />
Purification of gel slice of digested J61003.<br />
<br />
<br />
<center> [[image: Alberta_e1010DH5a_july12.jpg]] <br />
<br />
E1010 DH5a July 12</center><br />
<br />
-ED, JG, AL, JB, MC, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 13 ==<br />
<br />
'''General Notes:'''<br />
<br />
Everyone who is coming tonight please be present 7:00pm sharp<br />
<br />
'''Lab Notes:'''<br />
<br />
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.<br />
<br />
-ED, JB<br />
<br />
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"<br><br />
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)<br><br />
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL<br><br />
Made some Kan plates<br><br />
<br />
-MC, JP, JG, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 14 ==<br />
<b>Schedule:</b><br />
<br />
AL,JP,NK<br />
<br />
<b>General Notes:</b><br />
<br />
If you are scheduled during the day please be in the lab at 1230hrs (PM) <br />
Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911<br />
<br />
Things to be continued today (CZ)<br><br />
1. Put LB plates in 4 degree fridge <br><br />
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control<br><br />
3. Xba1/EcoRI double digest <br><br />
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour. <br><br />
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.<br><br />
<br />
If you need anything else call me(Celine). Number on the blackboard. Thank you.�<br />
<br />
<b/>Lab Notes:</b><br />
JP, AL, NK, JB(?) <br><br />
<br />
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20 <br><br />
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20 <br><br />
3. Placed Kan plates at +4 <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 15 ==<br />
<b>Schedule:</b><br />
<br />
Jp,JB,AL<br />
<br />
<b>General Notes:</b><br />
<br />
Options: <br><br />
<br />
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)<br><br />
2. gel purify J61003 restriction products (promoter restriction sample) <br><br />
3. fundraising letters! <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow<br><br />
2. glycerol stocks of Chlorobium tepidum<br><br />
3. finished grant application forms<br><br />
- JP, AL, NG, MC<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 16 ==<br />
<b>Schedule:</b><br />
<br />
CZ,VH,AF<br />
<br />
<b>General Notes:</b><br />
<br />
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP) <br><br />
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining <br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed gel extraction of E,X digested J61003. (in box in -20)<br />
<br />
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.<br />
<br />
-VH, ED, CZ<br />
<br />
<br />
'''Quotes of the Day:'''<br />
<br />
"If I had a remote, I'd change the cd."<br />
<br />
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it<br />
<br />
"I would never drive this tired. But I WILL operate chemicals."<br />
<br />
Anonymous<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 17 ==<br />
<b>Schedule:</b><br />
<br />
ED,VH,AF<br />
<br />
'''Lab Notes:'''<br />
<br />
Gel extracted E1010 from yesterday's slices.<br />
<br />
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post. <br><br />
<b> It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC </b> <br><br />
<br />
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED<br />
<br />
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.<br />
<br />
-ED,JG,VH,AF<br />
<br />
'''Quote of the Day'''<br />
<br />
"Cancer is an urban myth."<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,MC,NG<br />
<br />
<b>Lab Notes:</b><br />
<br />
Transformed Ligations 1,2,3,4 into XL10 Gold Competent cells <br><br />
Plated 1ul,10 ul, and 200ul on AMP <br><br />
Incubated @ 37celsius overnight<br><br />
<br />
-MC, NG, CZ<br />
<br />
<b>Quote(s) of the Day:</b><br />
<br />
"Asuuuuuuuuuuuum!!" <br><br />
In response to the awesome; "You're wayyyy to exciting for me right now" <br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 19 ==<br />
<br />
Meeting at 7:00, CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
ED,MC,JG<br />
<br />
<b>General Notes</b><br />
<br />
New Schedule is up<br><br />
check out the jar o' love @ the front :) <br><br />
<br />
<b>Lab notes</b><br />
<br />
Started 8X5mL overnights to test colonies of the RFP ligation. Hopefully JB will be available to do minipreps tomorrow. Miniprep protocol is tucked in Master Book.<br />
<br />
-ED,JG,MC<br />
<br />
<b>Quote of the Day </b><br />
<br />
"There is a pee throw up smell it the washroom"<br />
<br />
Reply<br />
<br />
"Someone must be pregnant"<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 20 ==<br />
<b>Schedule:</b><br />
<br />
JG,ED,MC <br><br />
<br />
<b>General Notes:</b><br><br />
<br />
There is now a Jar O' Love in the lab; Please do not drink chemicals from said jar o'love. Check it out - it's at the front on the instructors bench. It's to give shout outs for people who are rocking out and 'taking one for the team'. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprep Lig3 (8 samples) overnights - JB<br><br />
Made 1L LB broth (16 bottles) at the back to be put away in the sterile cupboards tomorrow <br><br />
Ran Gel of JB's Minipreps after digestion with Xba and Spe. All of the colonies were positive (band at 1.2 kB).<br><br />
Did some dishes - we should learn how to use the dishwasher<br><br />
<br />
- MC, ED, JG <br><br />
<br />
<b>Quote(s) of the day:</b><br><br />
<br />
"Always use protection when using the Jar O' Love"<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 21 ==<br />
<b>Schedule:</b><br />
<br />
JP,NK,AL<br />
<br />
<b>General Notes:</b><br />
<br />
<br />
Since it doesn't look like Nik will be showing up, maybe just call Alex and see if it works for him.<br />
<br />
All of the ligations of RFP into J61003 turned out. They are labelled JB July 19 1-8 in the DNA box. Transform 1 of them into XL10 Gold and plate on Amp with Tetracycline plates. You will have to add the Tet to the plates beforehand. I don't know what concentraion, but James suggested 1 ug/mL. Try plating different amounts.<br />
<br />
The reason that I am making these suggestions is that I read that XL10Golds are Tet resistant, so we can maybe induce the expression of RFP with the Tet because it is under the Tet promotor and then we can see the RFP.<br />
<br />
As always, call me if you need anything.<br />
<br />
-ED<br />
<br />
<b>Lab Notes:</b><br />
<br />
1. got into building <br><br />
2. waited for peers - no shows <br><br />
3. tried to break into room by climbing through roof (cement goes all the way up) <br><br />
4. went home... will complete work tomorrow - sorry i'll get swipe access asap... <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 22 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,AL,NG<br />
<br />
<b>General Notes:</b><br />
<br />
How does 7:00pm sound?<br />
<br />
<b>Laboratory Notes:</b><br />
<br />
Transformed XL10 gold cells with J61003 (with RFP ligated in). Split cell aliquot into two 50microL aliquots and added 1microL DNA from ligation sample 1 and from ligation sample 4. Covered AMP plates with 50microL 1microg/ml tet, spread, let sit. Plated (1) 50microL sample and (1) 5microL + 45microL LB onto Tet/Amp plates for #'s 1&4. They are in 37C incubator. The remaining transformed cells are in +4C labeled July 22. <br><br />
<br />
<b>Serious Quote of the Month</b><br />
<br />
<b>Faust</b><br><br />
Then shall I see, with vision clear, <br><br />
How secret elements cohere, <br><br />
And what the universe engirds, <br><br />
And give up huckstering with words <br><br />
<br />
-Johann Wolfgang von Goethe <br><br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,CZ,JG<br />
<br />
<b>Laboratory Notes:</b><br><br />
1. Add more Tet on ligation plates from July 22. Hopefully red colonies will appear!<br><br />
2. Transformation of XL10Gold with<br><br />
- Butanol CoA dehydrogenase (1 in 1000 dilution)<br><br />
- Butenoyl CoA dehydrogenase (1 in 1000 dilution)<br><br />
- J23018 (RFP positive control)<br><br />
<br />
20 and 100 microlitres of transformed culture plated on Amp plate and incubate at 37 degrees<br><br />
<br />
The leftover mixture is stored in 4 degree fridge in case of no growth.<br><br />
3. Repeate overnight ligation from July 22. The exact amounts of insert, vector etc. are on the blackboard.<br><br />
<br />
For tomorrow:<br><br />
1. Pick colonies for transformed plates.<br><br />
2. Plate ligation mixtures with appropriate amount of Tet.<br><br />
3. Observe red colonies from the July 22 ligation plates if the cells are still alive.<br><br />
<br />
Memorable Quote:<br><br />
Don't force it. Let it come out naturally.<br><br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,NG<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed I0500 (arabinose promotor) into HB101 competent cells. Plated 2, 20, 200 uL on Kan.<br />
<br />
Transformed ligations from yesterday into XL10 gold. Plated 100 uL on Amp.<br />
<br />
RFP control from yesterday worked, but sadly our ligations are not expressing any RFP. It was still exciting to see red control cells. It seems as if the Tet promotor is induced at a low level without any Tet.<br />
<br />
Started 4X5ml overnights of enoyl coA hydratase and butyryl coa DH. There seems to be some confusion with the naming of these proteins. These are the names that I have used in the BioBricks. We will be able to tell if they are correctly labelled when we do the digests. Enoyl coa hydratase is about 840 bp and butyryl coa DH is about 1200 bp.<br />
<br />
Note on agarose gels: Use only the wide combs (8 lanes) because we are having to many problems with the narrow lanes. This may mean that you will need to pour multiple gels (oh my). Also, try and run two ladders per gel until we get the ladder problem sorted out. Also, make sure that you zoom in on the gel when taking a picture.<br />
<br />
Note on Tet: should be stored 12.5 mg/mL in 50% ethanol, 50% water solution. This is a 1000X solution. Keep this in mind when using Tet again.<br />
<br />
-ED, AF, James<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Jp,MC,VH<br />
<br />
<b>General Notes:</b><br />
<br />
Use of wide combs- 5microL of sample works well with 3.5microL of ladder. Make sure you stir up the ladder before use!<br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed minipreps of enoylcoa hydratase and butyryl coa dh. They are in the new box with the green tape. I think that there are two types of digests we should be doing: one to make sure we have the right stuff (digest with EcoR1 and Pst1) and one to clone in the RBS. To clone in the RBS, cut B0034 with Eco and Spe and cut the enoyl coa hydratase and butyryl coa dh with Eco and Xba.<br />
<br />
Check to see if any colonies have grown on I0500 plates in incubator. There weren't any visible on Adam's transformations this morning. James also transformed some this morning, so they may be hard to see. If there are colonies, start overnights and see if Jori can do minipreps tomorrow. If Jori can't make it, get ahold of me and I can. AND if there are no colonies, just post here and I'll go check the plates in the morning.<br />
<br />
I also forgot to start glycerol stocks of our enoly coa hydratase and butyryl coa dh. If you guys could restart some overnights from the plates in the fridge so I can do it tomorrow that would be great.<br />
<br />
I had to rush to work this morning and didn't get any of this written in the lab book. So you guys can make fun of me or whatever. I guess I belong in the Science Hall of Shame.<br />
<br />
-ED <br><br />
<br />
Enny = Enoly CoA Hydratase<br><br />
Benny = Butytyl CoA DH<br />
<br />
Checked plates<br><br />
- Adam - no growth: parafilmed and put in 4 fridge<br><br />
- James'? J23018: red colonies parafilmed and put in 4 fridge (was this the control?)<br><br />
<br />
Overnights of Benny & Enny<br><br />
- 3x 5mlLB each <br><br />
<br />
Restrictions<br><br />
- B0034 with SPE & ECO<br><br />
- Enny with Xba & ECO<br><br />
-Benny with Xba & ECO<br><br />
These are stored in the -20 freezer<br><br />
<br />
- MC, NK, VH, NG, JP<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- Glycerol stocks of Enny & Benny (these colonies are likely different from the transformations as it was not marked out which colonies were picked for transformations)<br><br />
- Gels for restrictions of B0034, Enny & Benny<br />
- cut out the bands; gel extractions? (if there is time)<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 26 ==<br />
<br />
Meeting at 7:00, Room CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
AL,VH,ED<br />
<br />
<b>Lab Book</b><br />
<br />
Ran gel of "Enny", "Benny" and Boo IV against 1 kb ladder.<br />
<br />
Cleared up glasswares in the autoclave.<br />
<br />
Parafilm all unparafilmed plates.<br />
<br />
Checked out red fluorescence of J23018.<br />
<br />
Attempted working on fundraising letters (?)<br />
<br />
-ED, VH, AL, JG, MC, AF, Doug<br />
<br />
<b>Favourite "F" word of the day:</b><br />
<br />
fulcrum<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,MC,NK<br><br />
The theme is Foul music Friday - bring any bad music you have (or embarassed that you have in your collection)<br />
<br />
<b>General Notes:</b><br />
<br />
1. This is whats on the agenda for this evening:<br />
[[Image: Alberta_promoterprob.jpg]]<br />
<br />
<b>Lab book:</b><br />
<br />
Digest Enny(1,4), Benny (1,2) B0034(2,4) with PST, XBA, SPE<br><br />
Enny with PST and XBA<br><br />
Benny with PST and XBA<br><br />
B0034 with PST and SPE<br><br />
O/N of Benny and Enny glycerol stock (3 each)<br><br />
<br />
'''''Note the 4 degree fridge at the back is out of commision. All iGEM materials have been moved to the room where we take pictures of gels. The door labelled IGEM.''''' Nik is here on saturday and Celine can show on sunday.<br><br />
<br />
- JG, NK, MC, JP, and James (cos he loves us) <br />
<br />
<br />
<b>Jame's Fun Tip of the Day:</b><br />
If you want to run a gel for a shorter period of time ad the EtBR to the gel rather than the buffer. You can add ~4ul to the gel rather than 10ul to the buffer. <br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JB,NK<br />
<br />
<b>Meeting @ 1300hrs - MC on behalf of CZ</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Glycerol stocks of Enny & Benny from the O/N<br><br />
- Gels of Enny, Benny & B0034 (put the EtBr in the gel)<br><br />
- Ligations of Enny+Benny into B0034 with T4 ligase; leave O/N in incubator at the back room (near the phone) set at 13degrees celsius<br />
<br />
<b>Lab Book</b><br />
<br />
Finished making glycerol stocks of Enny and Benny from the O/N. Followed Erin's protocols except the Flash Freeze step.<br><br />
<br />
Ran agarose gel of Benny, Enny, b0034 digest.<br> <br />
No DnA ladder (NEVER DO IT AGAIN). <br><br />
Ran at 90V for 20 Min, run longer for next time.<br><br />
<br />
Cut Benny band (1200BP) and B0034, need to redigest Enny tomorrow<br><br />
Gel extration using QIAGEN kit<br><br />
<br />
Ligation , control only with ligase, <br><br />
Stored at 13 degrees celsius overnight <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,AF,NG<br><br />
<br />
<b>As agreed on Thursday meeting at noon or 12</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Transformation of ligation (Benny and B0034( into XL10 Gold <br> on Amp plates<br />
- Repeat double digest of nny, gel, and gel extration, and ligation<bR><br />
- Make AMP and KAN plates<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,JP,JG,<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 31 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,MC<br />
<br />
I cannot make it this day, as I will still be out of town. Can someone switch for Wednesday?<br />
-ED<br />
<br />
Whoops my Bad I can take it as long as me michelle and adam can handle the task without you justin or celine<br />
-JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/June|To June 2007]]<br><br />
[[Alberta/Calender/August|To August 2007]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/JulyAlberta/Calender/July2007-07-29T02:42:28Z<p>CelineYZ: /* July 29 */</p>
<hr />
<div><b>Schedule Legend:</B><br />
JG: Jason, <br />
Mc: Michelle,<br />
ED: Erin,<br />
NG: Nick G.,<br />
NK: Nik K.,<br />
CZ: Celine,<br />
AF: Adam,<br />
Al: Alex,<br />
JB: Jori,<br />
JP: Justin,<br />
VH: Veronica,<br />
<br />
<br />
=='''July'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<br />
<br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/July|July 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_1|1]]</td><br />
<td>[[Alberta/Calender/July#July_2|2]]</td><br />
<td>[[Alberta/Calender/July#July_3|3]]</td><br />
<td>[[Alberta/Calender/July#July_4|4]]</td><br />
<td>[[Alberta/Calender/July#July_5|5]]</td><br />
<td>[[Alberta/Calender/July#July_6|6]]</td><br />
<td>[[Alberta/Calender/July#July_7|7]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_8|8]]</td><br />
<td>[[Alberta/Calender/July#July_9|9]]</td><br />
<td>[[Alberta/Calender/July#July_10|10]]</td><br />
<td>[[Alberta/Calender/July#July_11|11]]</td><br />
<td>[[Alberta/Calender/July#July_12|12]]</td><br />
<td>[[Alberta/Calender/July#July_13|13]]</td><br />
<td>[[Alberta/Calender/July#July_14|14]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_15|15]]</td><br />
<td>[[Alberta/Calender/July#July_16|16]]</td><br />
<td>[[Alberta/Calender/July#July_17|17]]</td><br />
<td>[[Alberta/Calender/July#July_18|18]]</td><br />
<td>[[Alberta/Calender/July#July_19|19]]</td><br />
<td>[[Alberta/Calender/July#July_20|20]]</td><br />
<td>[[Alberta/Calender/July#July_21|21]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_22|22]]</td><br />
<td>[[Alberta/Calender/July#July_23|23]]</td><br />
<td>[[Alberta/Calender/July#July_24|24]]</td><br />
<td>[[Alberta/Calender/July#July_25|25]]</td><br />
<td>[[Alberta/Calender/July#July_26|26]]</td><br />
<td>[[Alberta/Calender/July#July_27|27]]</td><br />
<td>[[Alberta/Calender/July#July_28|28]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_29|29]]</td><br />
<td>[[Alberta/Calender/July#July_30|30]]</td><br />
<td>[[Alberta/Calender/July#July_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<br />
<br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== July 1 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 3 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 4 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.<br />
<br />
-ED, JG, ML, MC, VH, AL, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 6 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformation of E1010 did not work, it is in a Kan vector. SMRT.<br />
<br />
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 7 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.<br />
<br />
-JP,JB,AL<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 8 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.<br />
<br />
Pour Kan and Amp plates. Make TBE and TAE buffer.<br />
<br />
-ED, JP, MC, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 9 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.<br />
<br />
-ED, VH, JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 10 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.<br />
<br />
-ED, MC, AF, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 11 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.<br />
<br />
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.<br />
<br />
<center> [[image: Alberta_e1010xlgold_july11.jpg]] <br />
<br />
E1010 XLGOLD 10 July 11</center><br />
<br />
-ED, MC, NG, JG, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 12 ==<br />
<br />
'''General Notes:'''<br />
<br />
Calender up and running.<br />
<br />
Meeting tonight at 7:00 in CSC 2-49.<br />
<br />
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!!<br />
-Justin<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.<br />
<br />
Streak for single colonies of E1010, A0500, J61003.<br />
<br />
Purification of gel slice of digested J61003.<br />
<br />
<br />
<center> [[image: Alberta_e1010DH5a_july12.jpg]] <br />
<br />
E1010 DH5a July 12</center><br />
<br />
-ED, JG, AL, JB, MC, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 13 ==<br />
<br />
'''General Notes:'''<br />
<br />
Everyone who is coming tonight please be present 7:00pm sharp<br />
<br />
'''Lab Notes:'''<br />
<br />
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.<br />
<br />
-ED, JB<br />
<br />
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"<br><br />
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)<br><br />
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL<br><br />
Made some Kan plates<br><br />
<br />
-MC, JP, JG, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 14 ==<br />
<b>Schedule:</b><br />
<br />
AL,JP,NK<br />
<br />
<b>General Notes:</b><br />
<br />
If you are scheduled during the day please be in the lab at 1230hrs (PM) <br />
Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911<br />
<br />
Things to be continued today (CZ)<br><br />
1. Put LB plates in 4 degree fridge <br><br />
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control<br><br />
3. Xba1/EcoRI double digest <br><br />
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour. <br><br />
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.<br><br />
<br />
If you need anything else call me(Celine). Number on the blackboard. Thank you.�<br />
<br />
<b/>Lab Notes:</b><br />
JP, AL, NK, JB(?) <br><br />
<br />
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20 <br><br />
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20 <br><br />
3. Placed Kan plates at +4 <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 15 ==<br />
<b>Schedule:</b><br />
<br />
Jp,JB,AL<br />
<br />
<b>General Notes:</b><br />
<br />
Options: <br><br />
<br />
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)<br><br />
2. gel purify J61003 restriction products (promoter restriction sample) <br><br />
3. fundraising letters! <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow<br><br />
2. glycerol stocks of Chlorobium tepidum<br><br />
3. finished grant application forms<br><br />
- JP, AL, NG, MC<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 16 ==<br />
<b>Schedule:</b><br />
<br />
CZ,VH,AF<br />
<br />
<b>General Notes:</b><br />
<br />
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP) <br><br />
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining <br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed gel extraction of E,X digested J61003. (in box in -20)<br />
<br />
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.<br />
<br />
-VH, ED, CZ<br />
<br />
<br />
'''Quotes of the Day:'''<br />
<br />
"If I had a remote, I'd change the cd."<br />
<br />
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it<br />
<br />
"I would never drive this tired. But I WILL operate chemicals."<br />
<br />
Anonymous<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 17 ==<br />
<b>Schedule:</b><br />
<br />
ED,VH,AF<br />
<br />
'''Lab Notes:'''<br />
<br />
Gel extracted E1010 from yesterday's slices.<br />
<br />
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post. <br><br />
<b> It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC </b> <br><br />
<br />
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED<br />
<br />
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.<br />
<br />
-ED,JG,VH,AF<br />
<br />
'''Quote of the Day'''<br />
<br />
"Cancer is an urban myth."<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,MC,NG<br />
<br />
<b>Lab Notes:</b><br />
<br />
Transformed Ligations 1,2,3,4 into XL10 Gold Competent cells <br><br />
Plated 1ul,10 ul, and 200ul on AMP <br><br />
Incubated @ 37celsius overnight<br><br />
<br />
-MC, NG, CZ<br />
<br />
<b>Quote(s) of the Day:</b><br />
<br />
"Asuuuuuuuuuuuum!!" <br><br />
In response to the awesome; "You're wayyyy to exciting for me right now" <br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 19 ==<br />
<br />
Meeting at 7:00, CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
ED,MC,JG<br />
<br />
<b>General Notes</b><br />
<br />
New Schedule is up<br><br />
check out the jar o' love @ the front :) <br><br />
<br />
<b>Lab notes</b><br />
<br />
Started 8X5mL overnights to test colonies of the RFP ligation. Hopefully JB will be available to do minipreps tomorrow. Miniprep protocol is tucked in Master Book.<br />
<br />
-ED,JG,MC<br />
<br />
<b>Quote of the Day </b><br />
<br />
"There is a pee throw up smell it the washroom"<br />
<br />
Reply<br />
<br />
"Someone must be pregnant"<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 20 ==<br />
<b>Schedule:</b><br />
<br />
JG,ED,MC <br><br />
<br />
<b>General Notes:</b><br><br />
<br />
There is now a Jar O' Love in the lab; Please do not drink chemicals from said jar o'love. Check it out - it's at the front on the instructors bench. It's to give shout outs for people who are rocking out and 'taking one for the team'. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprep Lig3 (8 samples) overnights - JB<br><br />
Made 1L LB broth (16 bottles) at the back to be put away in the sterile cupboards tomorrow <br><br />
Ran Gel of JB's Minipreps after digestion with Xba and Spe. All of the colonies were positive (band at 1.2 kB).<br><br />
Did some dishes - we should learn how to use the dishwasher<br><br />
<br />
- MC, ED, JG <br><br />
<br />
<b>Quote(s) of the day:</b><br><br />
<br />
"Always use protection when using the Jar O' Love"<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 21 ==<br />
<b>Schedule:</b><br />
<br />
JP,NK,AL<br />
<br />
<b>General Notes:</b><br />
<br />
<br />
Since it doesn't look like Nik will be showing up, maybe just call Alex and see if it works for him.<br />
<br />
All of the ligations of RFP into J61003 turned out. They are labelled JB July 19 1-8 in the DNA box. Transform 1 of them into XL10 Gold and plate on Amp with Tetracycline plates. You will have to add the Tet to the plates beforehand. I don't know what concentraion, but James suggested 1 ug/mL. Try plating different amounts.<br />
<br />
The reason that I am making these suggestions is that I read that XL10Golds are Tet resistant, so we can maybe induce the expression of RFP with the Tet because it is under the Tet promotor and then we can see the RFP.<br />
<br />
As always, call me if you need anything.<br />
<br />
-ED<br />
<br />
<b>Lab Notes:</b><br />
<br />
1. got into building <br><br />
2. waited for peers - no shows <br><br />
3. tried to break into room by climbing through roof (cement goes all the way up) <br><br />
4. went home... will complete work tomorrow - sorry i'll get swipe access asap... <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 22 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,AL,NG<br />
<br />
<b>General Notes:</b><br />
<br />
How does 7:00pm sound?<br />
<br />
<b>Laboratory Notes:</b><br />
<br />
Transformed XL10 gold cells with J61003 (with RFP ligated in). Split cell aliquot into two 50microL aliquots and added 1microL DNA from ligation sample 1 and from ligation sample 4. Covered AMP plates with 50microL 1microg/ml tet, spread, let sit. Plated (1) 50microL sample and (1) 5microL + 45microL LB onto Tet/Amp plates for #'s 1&4. They are in 37C incubator. The remaining transformed cells are in +4C labeled July 22. <br><br />
<br />
<b>Serious Quote of the Month</b><br />
<br />
<b>Faust</b><br><br />
Then shall I see, with vision clear, <br><br />
How secret elements cohere, <br><br />
And what the universe engirds, <br><br />
And give up huckstering with words <br><br />
<br />
-Johann Wolfgang von Goethe <br><br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,CZ,JG<br />
<br />
<b>Laboratory Notes:</b><br><br />
1. Add more Tet on ligation plates from July 22. Hopefully red colonies will appear!<br><br />
2. Transformation of XL10Gold with<br><br />
- Butanol CoA dehydrogenase (1 in 1000 dilution)<br><br />
- Butenoyl CoA dehydrogenase (1 in 1000 dilution)<br><br />
- J23018 (RFP positive control)<br><br />
<br />
20 and 100 microlitres of transformed culture plated on Amp plate and incubate at 37 degrees<br><br />
<br />
The leftover mixture is stored in 4 degree fridge in case of no growth.<br><br />
3. Repeate overnight ligation from July 22. The exact amounts of insert, vector etc. are on the blackboard.<br><br />
<br />
For tomorrow:<br><br />
1. Pick colonies for transformed plates.<br><br />
2. Plate ligation mixtures with appropriate amount of Tet.<br><br />
3. Observe red colonies from the July 22 ligation plates if the cells are still alive.<br><br />
<br />
Memorable Quote:<br><br />
Don't force it. Let it come out naturally.<br><br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,NG<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed I0500 (arabinose promotor) into HB101 competent cells. Plated 2, 20, 200 uL on Kan.<br />
<br />
Transformed ligations from yesterday into XL10 gold. Plated 100 uL on Amp.<br />
<br />
RFP control from yesterday worked, but sadly our ligations are not expressing any RFP. It was still exciting to see red control cells. It seems as if the Tet promotor is induced at a low level without any Tet.<br />
<br />
Started 4X5ml overnights of enoyl coA hydratase and butyryl coa DH. There seems to be some confusion with the naming of these proteins. These are the names that I have used in the BioBricks. We will be able to tell if they are correctly labelled when we do the digests. Enoyl coa hydratase is about 840 bp and butyryl coa DH is about 1200 bp.<br />
<br />
Note on agarose gels: Use only the wide combs (8 lanes) because we are having to many problems with the narrow lanes. This may mean that you will need to pour multiple gels (oh my). Also, try and run two ladders per gel until we get the ladder problem sorted out. Also, make sure that you zoom in on the gel when taking a picture.<br />
<br />
Note on Tet: should be stored 12.5 mg/mL in 50% ethanol, 50% water solution. This is a 1000X solution. Keep this in mind when using Tet again.<br />
<br />
-ED, AF, James<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Jp,MC,VH<br />
<br />
<b>General Notes:</b><br />
<br />
Use of wide combs- 5microL of sample works well with 3.5microL of ladder. Make sure you stir up the ladder before use!<br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed minipreps of enoylcoa hydratase and butyryl coa dh. They are in the new box with the green tape. I think that there are two types of digests we should be doing: one to make sure we have the right stuff (digest with EcoR1 and Pst1) and one to clone in the RBS. To clone in the RBS, cut B0034 with Eco and Spe and cut the enoyl coa hydratase and butyryl coa dh with Eco and Xba.<br />
<br />
Check to see if any colonies have grown on I0500 plates in incubator. There weren't any visible on Adam's transformations this morning. James also transformed some this morning, so they may be hard to see. If there are colonies, start overnights and see if Jori can do minipreps tomorrow. If Jori can't make it, get ahold of me and I can. AND if there are no colonies, just post here and I'll go check the plates in the morning.<br />
<br />
I also forgot to start glycerol stocks of our enoly coa hydratase and butyryl coa dh. If you guys could restart some overnights from the plates in the fridge so I can do it tomorrow that would be great.<br />
<br />
I had to rush to work this morning and didn't get any of this written in the lab book. So you guys can make fun of me or whatever. I guess I belong in the Science Hall of Shame.<br />
<br />
-ED <br><br />
<br />
Enny = Enoly CoA Hydratase<br><br />
Benny = Butytyl CoA DH<br />
<br />
Checked plates<br><br />
- Adam - no growth: parafilmed and put in 4 fridge<br><br />
- James'? J23018: red colonies parafilmed and put in 4 fridge (was this the control?)<br><br />
<br />
Overnights of Benny & Enny<br><br />
- 3x 5mlLB each <br><br />
<br />
Restrictions<br><br />
- B0034 with SPE & ECO<br><br />
- Enny with Xba & ECO<br><br />
-Benny with Xba & ECO<br><br />
These are stored in the -20 freezer<br><br />
<br />
- MC, NK, VH, NG, JP<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- Glycerol stocks of Enny & Benny (these colonies are likely different from the transformations as it was not marked out which colonies were picked for transformations)<br><br />
- Gels for restrictions of B0034, Enny & Benny<br />
- cut out the bands; gel extractions? (if there is time)<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 26 ==<br />
<br />
Meeting at 7:00, Room CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
AL,VH,ED<br />
<br />
<b>Lab Book</b><br />
<br />
Ran gel of "Enny", "Benny" and Boo IV against 1 kb ladder.<br />
<br />
Cleared up glasswares in the autoclave.<br />
<br />
Parafilm all unparafilmed plates.<br />
<br />
Checked out red fluorescence of J23018.<br />
<br />
Attempted working on fundraising letters (?)<br />
<br />
-ED, VH, AL, JG, MC, AF, Doug<br />
<br />
<b>Favourite "F" word of the day:</b><br />
<br />
fulcrum<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,MC,NK<br><br />
The theme is Foul music Friday - bring any bad music you have (or embarassed that you have in your collection)<br />
<br />
<b>General Notes:</b><br />
<br />
1. This is whats on the agenda for this evening:<br />
[[Image: Alberta_promoterprob.jpg]]<br />
<br />
<b>Lab book:</b><br />
<br />
Digest Enny(1,4), Benny (1,2) B0034(2,4) with PST, XBA, SPE<br><br />
Enny with PST and XBA<br><br />
Benny with PST and XBA<br><br />
B0034 with PST and SPE<br><br />
O/N of Benny and Enny glycerol stock (3 each)<br><br />
<br />
'''''Note the 4 degree fridge at the back is out of commision. All iGEM materials have been moved to the room where we take pictures of gels. The door labelled IGEM.''''' Nik is here on saturday and Celine can show on sunday.<br><br />
<br />
- JG, NK, MC, JP, and James (cos he loves us) <br />
<br />
<br />
<b>Jame's Fun Tip of the Day:</b><br />
If you want to run a gel for a shorter period of time ad the EtBR to the gel rather than the buffer. You can add ~4ul to the gel rather than 10ul to the buffer. <br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JB,NK<br />
<br />
<b>Meeting @ 1300hrs - MC on behalf of CZ</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Glycerol stocks of Enny & Benny from the O/N<br><br />
- Gels of Enny, Benny & B0034 (put the EtBr in the gel)<br><br />
- Ligations of Enny+Benny into B0034 with T4 ligase; leave O/N in incubator at the back room (near the phone) set at 13degrees celsius<br />
<br />
<b>Lab Book</b><br />
<br />
Finished making glycerol stocks of Enny and Benny from the O/N. Followed Erin's protocols except the Flash Freeze step.<br><br />
<br />
Ran agarose gel of Benny, Enny, b0034 digest.<br> <br />
No DnA ladder (NEVER DO IT AGAIN). <br><br />
Ran at 90V for 20 Min, run longer for next time.<br><br />
<br />
Cut Benny band (1200BP) and B0034, need to redigest Enny tomorrow<br><br />
Gel extration using QIAGEN kit<br><br />
<br />
Ligation , control only with ligase, <br><br />
Stored at 13 degrees celsius overnight <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,AF,NG<br><br />
<br />
<b>As agreed on Thursday meeting at noon or 12</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Transformation of ligation (Benny and B0034( into XL10 Gold <br> on Amp plates<br />
- Repeat double digest of nny, gel, and gel extration, and ligation<bR><br />
- Make AMP and KAM plates<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,JP,JG,<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 31 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,MC<br />
<br />
I cannot make it this day, as I will still be out of town. Can someone switch for Wednesday?<br />
-ED<br />
<br />
Whoops my Bad I can take it as long as me michelle and adam can handle the task without you justin or celine<br />
-JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/June|To June 2007]]<br><br />
[[Alberta/Calender/August|To August 2007]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/JulyAlberta/Calender/July2007-07-29T02:42:18Z<p>CelineYZ: /* July 29 */</p>
<hr />
<div><b>Schedule Legend:</B><br />
JG: Jason, <br />
Mc: Michelle,<br />
ED: Erin,<br />
NG: Nick G.,<br />
NK: Nik K.,<br />
CZ: Celine,<br />
AF: Adam,<br />
Al: Alex,<br />
JB: Jori,<br />
JP: Justin,<br />
VH: Veronica,<br />
<br />
<br />
=='''July'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<br />
<br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/July|July 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_1|1]]</td><br />
<td>[[Alberta/Calender/July#July_2|2]]</td><br />
<td>[[Alberta/Calender/July#July_3|3]]</td><br />
<td>[[Alberta/Calender/July#July_4|4]]</td><br />
<td>[[Alberta/Calender/July#July_5|5]]</td><br />
<td>[[Alberta/Calender/July#July_6|6]]</td><br />
<td>[[Alberta/Calender/July#July_7|7]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_8|8]]</td><br />
<td>[[Alberta/Calender/July#July_9|9]]</td><br />
<td>[[Alberta/Calender/July#July_10|10]]</td><br />
<td>[[Alberta/Calender/July#July_11|11]]</td><br />
<td>[[Alberta/Calender/July#July_12|12]]</td><br />
<td>[[Alberta/Calender/July#July_13|13]]</td><br />
<td>[[Alberta/Calender/July#July_14|14]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_15|15]]</td><br />
<td>[[Alberta/Calender/July#July_16|16]]</td><br />
<td>[[Alberta/Calender/July#July_17|17]]</td><br />
<td>[[Alberta/Calender/July#July_18|18]]</td><br />
<td>[[Alberta/Calender/July#July_19|19]]</td><br />
<td>[[Alberta/Calender/July#July_20|20]]</td><br />
<td>[[Alberta/Calender/July#July_21|21]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_22|22]]</td><br />
<td>[[Alberta/Calender/July#July_23|23]]</td><br />
<td>[[Alberta/Calender/July#July_24|24]]</td><br />
<td>[[Alberta/Calender/July#July_25|25]]</td><br />
<td>[[Alberta/Calender/July#July_26|26]]</td><br />
<td>[[Alberta/Calender/July#July_27|27]]</td><br />
<td>[[Alberta/Calender/July#July_28|28]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_29|29]]</td><br />
<td>[[Alberta/Calender/July#July_30|30]]</td><br />
<td>[[Alberta/Calender/July#July_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<br />
<br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== July 1 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 3 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 4 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.<br />
<br />
-ED, JG, ML, MC, VH, AL, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 6 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformation of E1010 did not work, it is in a Kan vector. SMRT.<br />
<br />
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 7 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.<br />
<br />
-JP,JB,AL<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 8 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.<br />
<br />
Pour Kan and Amp plates. Make TBE and TAE buffer.<br />
<br />
-ED, JP, MC, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 9 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.<br />
<br />
-ED, VH, JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 10 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.<br />
<br />
-ED, MC, AF, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 11 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.<br />
<br />
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.<br />
<br />
<center> [[image: Alberta_e1010xlgold_july11.jpg]] <br />
<br />
E1010 XLGOLD 10 July 11</center><br />
<br />
-ED, MC, NG, JG, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 12 ==<br />
<br />
'''General Notes:'''<br />
<br />
Calender up and running.<br />
<br />
Meeting tonight at 7:00 in CSC 2-49.<br />
<br />
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!!<br />
-Justin<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.<br />
<br />
Streak for single colonies of E1010, A0500, J61003.<br />
<br />
Purification of gel slice of digested J61003.<br />
<br />
<br />
<center> [[image: Alberta_e1010DH5a_july12.jpg]] <br />
<br />
E1010 DH5a July 12</center><br />
<br />
-ED, JG, AL, JB, MC, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 13 ==<br />
<br />
'''General Notes:'''<br />
<br />
Everyone who is coming tonight please be present 7:00pm sharp<br />
<br />
'''Lab Notes:'''<br />
<br />
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.<br />
<br />
-ED, JB<br />
<br />
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"<br><br />
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)<br><br />
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL<br><br />
Made some Kan plates<br><br />
<br />
-MC, JP, JG, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 14 ==<br />
<b>Schedule:</b><br />
<br />
AL,JP,NK<br />
<br />
<b>General Notes:</b><br />
<br />
If you are scheduled during the day please be in the lab at 1230hrs (PM) <br />
Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911<br />
<br />
Things to be continued today (CZ)<br><br />
1. Put LB plates in 4 degree fridge <br><br />
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control<br><br />
3. Xba1/EcoRI double digest <br><br />
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour. <br><br />
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.<br><br />
<br />
If you need anything else call me(Celine). Number on the blackboard. Thank you.�<br />
<br />
<b/>Lab Notes:</b><br />
JP, AL, NK, JB(?) <br><br />
<br />
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20 <br><br />
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20 <br><br />
3. Placed Kan plates at +4 <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 15 ==<br />
<b>Schedule:</b><br />
<br />
Jp,JB,AL<br />
<br />
<b>General Notes:</b><br />
<br />
Options: <br><br />
<br />
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)<br><br />
2. gel purify J61003 restriction products (promoter restriction sample) <br><br />
3. fundraising letters! <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow<br><br />
2. glycerol stocks of Chlorobium tepidum<br><br />
3. finished grant application forms<br><br />
- JP, AL, NG, MC<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 16 ==<br />
<b>Schedule:</b><br />
<br />
CZ,VH,AF<br />
<br />
<b>General Notes:</b><br />
<br />
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP) <br><br />
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining <br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed gel extraction of E,X digested J61003. (in box in -20)<br />
<br />
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.<br />
<br />
-VH, ED, CZ<br />
<br />
<br />
'''Quotes of the Day:'''<br />
<br />
"If I had a remote, I'd change the cd."<br />
<br />
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it<br />
<br />
"I would never drive this tired. But I WILL operate chemicals."<br />
<br />
Anonymous<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 17 ==<br />
<b>Schedule:</b><br />
<br />
ED,VH,AF<br />
<br />
'''Lab Notes:'''<br />
<br />
Gel extracted E1010 from yesterday's slices.<br />
<br />
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post. <br><br />
<b> It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC </b> <br><br />
<br />
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED<br />
<br />
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.<br />
<br />
-ED,JG,VH,AF<br />
<br />
'''Quote of the Day'''<br />
<br />
"Cancer is an urban myth."<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,MC,NG<br />
<br />
<b>Lab Notes:</b><br />
<br />
Transformed Ligations 1,2,3,4 into XL10 Gold Competent cells <br><br />
Plated 1ul,10 ul, and 200ul on AMP <br><br />
Incubated @ 37celsius overnight<br><br />
<br />
-MC, NG, CZ<br />
<br />
<b>Quote(s) of the Day:</b><br />
<br />
"Asuuuuuuuuuuuum!!" <br><br />
In response to the awesome; "You're wayyyy to exciting for me right now" <br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 19 ==<br />
<br />
Meeting at 7:00, CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
ED,MC,JG<br />
<br />
<b>General Notes</b><br />
<br />
New Schedule is up<br><br />
check out the jar o' love @ the front :) <br><br />
<br />
<b>Lab notes</b><br />
<br />
Started 8X5mL overnights to test colonies of the RFP ligation. Hopefully JB will be available to do minipreps tomorrow. Miniprep protocol is tucked in Master Book.<br />
<br />
-ED,JG,MC<br />
<br />
<b>Quote of the Day </b><br />
<br />
"There is a pee throw up smell it the washroom"<br />
<br />
Reply<br />
<br />
"Someone must be pregnant"<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 20 ==<br />
<b>Schedule:</b><br />
<br />
JG,ED,MC <br><br />
<br />
<b>General Notes:</b><br><br />
<br />
There is now a Jar O' Love in the lab; Please do not drink chemicals from said jar o'love. Check it out - it's at the front on the instructors bench. It's to give shout outs for people who are rocking out and 'taking one for the team'. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprep Lig3 (8 samples) overnights - JB<br><br />
Made 1L LB broth (16 bottles) at the back to be put away in the sterile cupboards tomorrow <br><br />
Ran Gel of JB's Minipreps after digestion with Xba and Spe. All of the colonies were positive (band at 1.2 kB).<br><br />
Did some dishes - we should learn how to use the dishwasher<br><br />
<br />
- MC, ED, JG <br><br />
<br />
<b>Quote(s) of the day:</b><br><br />
<br />
"Always use protection when using the Jar O' Love"<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 21 ==<br />
<b>Schedule:</b><br />
<br />
JP,NK,AL<br />
<br />
<b>General Notes:</b><br />
<br />
<br />
Since it doesn't look like Nik will be showing up, maybe just call Alex and see if it works for him.<br />
<br />
All of the ligations of RFP into J61003 turned out. They are labelled JB July 19 1-8 in the DNA box. Transform 1 of them into XL10 Gold and plate on Amp with Tetracycline plates. You will have to add the Tet to the plates beforehand. I don't know what concentraion, but James suggested 1 ug/mL. Try plating different amounts.<br />
<br />
The reason that I am making these suggestions is that I read that XL10Golds are Tet resistant, so we can maybe induce the expression of RFP with the Tet because it is under the Tet promotor and then we can see the RFP.<br />
<br />
As always, call me if you need anything.<br />
<br />
-ED<br />
<br />
<b>Lab Notes:</b><br />
<br />
1. got into building <br><br />
2. waited for peers - no shows <br><br />
3. tried to break into room by climbing through roof (cement goes all the way up) <br><br />
4. went home... will complete work tomorrow - sorry i'll get swipe access asap... <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 22 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,AL,NG<br />
<br />
<b>General Notes:</b><br />
<br />
How does 7:00pm sound?<br />
<br />
<b>Laboratory Notes:</b><br />
<br />
Transformed XL10 gold cells with J61003 (with RFP ligated in). Split cell aliquot into two 50microL aliquots and added 1microL DNA from ligation sample 1 and from ligation sample 4. Covered AMP plates with 50microL 1microg/ml tet, spread, let sit. Plated (1) 50microL sample and (1) 5microL + 45microL LB onto Tet/Amp plates for #'s 1&4. They are in 37C incubator. The remaining transformed cells are in +4C labeled July 22. <br><br />
<br />
<b>Serious Quote of the Month</b><br />
<br />
<b>Faust</b><br><br />
Then shall I see, with vision clear, <br><br />
How secret elements cohere, <br><br />
And what the universe engirds, <br><br />
And give up huckstering with words <br><br />
<br />
-Johann Wolfgang von Goethe <br><br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,CZ,JG<br />
<br />
<b>Laboratory Notes:</b><br><br />
1. Add more Tet on ligation plates from July 22. Hopefully red colonies will appear!<br><br />
2. Transformation of XL10Gold with<br><br />
- Butanol CoA dehydrogenase (1 in 1000 dilution)<br><br />
- Butenoyl CoA dehydrogenase (1 in 1000 dilution)<br><br />
- J23018 (RFP positive control)<br><br />
<br />
20 and 100 microlitres of transformed culture plated on Amp plate and incubate at 37 degrees<br><br />
<br />
The leftover mixture is stored in 4 degree fridge in case of no growth.<br><br />
3. Repeate overnight ligation from July 22. The exact amounts of insert, vector etc. are on the blackboard.<br><br />
<br />
For tomorrow:<br><br />
1. Pick colonies for transformed plates.<br><br />
2. Plate ligation mixtures with appropriate amount of Tet.<br><br />
3. Observe red colonies from the July 22 ligation plates if the cells are still alive.<br><br />
<br />
Memorable Quote:<br><br />
Don't force it. Let it come out naturally.<br><br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,NG<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed I0500 (arabinose promotor) into HB101 competent cells. Plated 2, 20, 200 uL on Kan.<br />
<br />
Transformed ligations from yesterday into XL10 gold. Plated 100 uL on Amp.<br />
<br />
RFP control from yesterday worked, but sadly our ligations are not expressing any RFP. It was still exciting to see red control cells. It seems as if the Tet promotor is induced at a low level without any Tet.<br />
<br />
Started 4X5ml overnights of enoyl coA hydratase and butyryl coa DH. There seems to be some confusion with the naming of these proteins. These are the names that I have used in the BioBricks. We will be able to tell if they are correctly labelled when we do the digests. Enoyl coa hydratase is about 840 bp and butyryl coa DH is about 1200 bp.<br />
<br />
Note on agarose gels: Use only the wide combs (8 lanes) because we are having to many problems with the narrow lanes. This may mean that you will need to pour multiple gels (oh my). Also, try and run two ladders per gel until we get the ladder problem sorted out. Also, make sure that you zoom in on the gel when taking a picture.<br />
<br />
Note on Tet: should be stored 12.5 mg/mL in 50% ethanol, 50% water solution. This is a 1000X solution. Keep this in mind when using Tet again.<br />
<br />
-ED, AF, James<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Jp,MC,VH<br />
<br />
<b>General Notes:</b><br />
<br />
Use of wide combs- 5microL of sample works well with 3.5microL of ladder. Make sure you stir up the ladder before use!<br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed minipreps of enoylcoa hydratase and butyryl coa dh. They are in the new box with the green tape. I think that there are two types of digests we should be doing: one to make sure we have the right stuff (digest with EcoR1 and Pst1) and one to clone in the RBS. To clone in the RBS, cut B0034 with Eco and Spe and cut the enoyl coa hydratase and butyryl coa dh with Eco and Xba.<br />
<br />
Check to see if any colonies have grown on I0500 plates in incubator. There weren't any visible on Adam's transformations this morning. James also transformed some this morning, so they may be hard to see. If there are colonies, start overnights and see if Jori can do minipreps tomorrow. If Jori can't make it, get ahold of me and I can. AND if there are no colonies, just post here and I'll go check the plates in the morning.<br />
<br />
I also forgot to start glycerol stocks of our enoly coa hydratase and butyryl coa dh. If you guys could restart some overnights from the plates in the fridge so I can do it tomorrow that would be great.<br />
<br />
I had to rush to work this morning and didn't get any of this written in the lab book. So you guys can make fun of me or whatever. I guess I belong in the Science Hall of Shame.<br />
<br />
-ED <br><br />
<br />
Enny = Enoly CoA Hydratase<br><br />
Benny = Butytyl CoA DH<br />
<br />
Checked plates<br><br />
- Adam - no growth: parafilmed and put in 4 fridge<br><br />
- James'? J23018: red colonies parafilmed and put in 4 fridge (was this the control?)<br><br />
<br />
Overnights of Benny & Enny<br><br />
- 3x 5mlLB each <br><br />
<br />
Restrictions<br><br />
- B0034 with SPE & ECO<br><br />
- Enny with Xba & ECO<br><br />
-Benny with Xba & ECO<br><br />
These are stored in the -20 freezer<br><br />
<br />
- MC, NK, VH, NG, JP<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- Glycerol stocks of Enny & Benny (these colonies are likely different from the transformations as it was not marked out which colonies were picked for transformations)<br><br />
- Gels for restrictions of B0034, Enny & Benny<br />
- cut out the bands; gel extractions? (if there is time)<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 26 ==<br />
<br />
Meeting at 7:00, Room CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
AL,VH,ED<br />
<br />
<b>Lab Book</b><br />
<br />
Ran gel of "Enny", "Benny" and Boo IV against 1 kb ladder.<br />
<br />
Cleared up glasswares in the autoclave.<br />
<br />
Parafilm all unparafilmed plates.<br />
<br />
Checked out red fluorescence of J23018.<br />
<br />
Attempted working on fundraising letters (?)<br />
<br />
-ED, VH, AL, JG, MC, AF, Doug<br />
<br />
<b>Favourite "F" word of the day:</b><br />
<br />
fulcrum<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,MC,NK<br><br />
The theme is Foul music Friday - bring any bad music you have (or embarassed that you have in your collection)<br />
<br />
<b>General Notes:</b><br />
<br />
1. This is whats on the agenda for this evening:<br />
[[Image: Alberta_promoterprob.jpg]]<br />
<br />
<b>Lab book:</b><br />
<br />
Digest Enny(1,4), Benny (1,2) B0034(2,4) with PST, XBA, SPE<br><br />
Enny with PST and XBA<br><br />
Benny with PST and XBA<br><br />
B0034 with PST and SPE<br><br />
O/N of Benny and Enny glycerol stock (3 each)<br><br />
<br />
'''''Note the 4 degree fridge at the back is out of commision. All iGEM materials have been moved to the room where we take pictures of gels. The door labelled IGEM.''''' Nik is here on saturday and Celine can show on sunday.<br><br />
<br />
- JG, NK, MC, JP, and James (cos he loves us) <br />
<br />
<br />
<b>Jame's Fun Tip of the Day:</b><br />
If you want to run a gel for a shorter period of time ad the EtBR to the gel rather than the buffer. You can add ~4ul to the gel rather than 10ul to the buffer. <br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JB,NK<br />
<br />
<b>Meeting @ 1300hrs - MC on behalf of CZ</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Glycerol stocks of Enny & Benny from the O/N<br><br />
- Gels of Enny, Benny & B0034 (put the EtBr in the gel)<br><br />
- Ligations of Enny+Benny into B0034 with T4 ligase; leave O/N in incubator at the back room (near the phone) set at 13degrees celsius<br />
<br />
<b>Lab Book</b><br />
<br />
Finished making glycerol stocks of Enny and Benny from the O/N. Followed Erin's protocols except the Flash Freeze step.<br><br />
<br />
Ran agarose gel of Benny, Enny, b0034 digest.<br> <br />
No DnA ladder (NEVER DO IT AGAIN). <br><br />
Ran at 90V for 20 Min, run longer for next time.<br><br />
<br />
Cut Benny band (1200BP) and B0034, need to redigest Enny tomorrow<br><br />
Gel extration using QIAGEN kit<br><br />
<br />
Ligation , control only with ligase, <br><br />
Stored at 13 degrees celsius overnight <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,AF,NG<br><br />
<br />
<b>As agreed on Thursday meeting at noon or 12</b><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Transformation of ligation (Benny and B0034( into XL10 Gold <br> on Amp plates<br />
- Repeat double digest of nny, gel, and gel extration, and ligation<bR><br />
- Make AMP and KAn plates<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,JP,JG,<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 31 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,MC<br />
<br />
I cannot make it this day, as I will still be out of town. Can someone switch for Wednesday?<br />
-ED<br />
<br />
Whoops my Bad I can take it as long as me michelle and adam can handle the task without you justin or celine<br />
-JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/June|To June 2007]]<br><br />
[[Alberta/Calender/August|To August 2007]]</div>CelineYZhttp://2007.igem.org/wiki/index.php/Alberta/Calender/JulyAlberta/Calender/July2007-07-28T07:42:24Z<p>CelineYZ: /* July 28 */</p>
<hr />
<div><b>Schedule Legend:</B><br />
JG: Jason, <br />
Mc: Michelle,<br />
ED: Erin,<br />
NG: Nick G.,<br />
NK: Nik K.,<br />
CZ: Celine,<br />
AF: Adam,<br />
Al: Alex,<br />
JB: Jori,<br />
JP: Justin,<br />
VH: Veronica,<br />
<br />
<br />
=='''July'''==<br />
<br />
<div valign="center"> <br />
<table><tr><td><table style="<br />
font-family: Verdana, Arial, Helvetica, sans-serif;<br />
vertical-align:middle;<br />
text-align:center;<br />
background-color:white;<br />
border-color:#FFFFFF;<br />
border-width:1px;<br />
color:black;<br />
font-size: 12px;"><br />
<tr><br />
<br />
<br />
<td colspan="7" style="font-weight: bold;<br />
width:170px;<br />
height:24px;<br />
color: black;">[[Alberta/Calender/July|July 2007]]</td><br />
</tr><br />
<tr><br />
<td>Su</td><br />
<td>M</td><br />
<td>Tu</td><br />
<td>W</td><br />
<td>Th</td><br />
<td>F</td><br />
<td>Sa</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_1|1]]</td><br />
<td>[[Alberta/Calender/July#July_2|2]]</td><br />
<td>[[Alberta/Calender/July#July_3|3]]</td><br />
<td>[[Alberta/Calender/July#July_4|4]]</td><br />
<td>[[Alberta/Calender/July#July_5|5]]</td><br />
<td>[[Alberta/Calender/July#July_6|6]]</td><br />
<td>[[Alberta/Calender/July#July_7|7]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_8|8]]</td><br />
<td>[[Alberta/Calender/July#July_9|9]]</td><br />
<td>[[Alberta/Calender/July#July_10|10]]</td><br />
<td>[[Alberta/Calender/July#July_11|11]]</td><br />
<td>[[Alberta/Calender/July#July_12|12]]</td><br />
<td>[[Alberta/Calender/July#July_13|13]]</td><br />
<td>[[Alberta/Calender/July#July_14|14]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_15|15]]</td><br />
<td>[[Alberta/Calender/July#July_16|16]]</td><br />
<td>[[Alberta/Calender/July#July_17|17]]</td><br />
<td>[[Alberta/Calender/July#July_18|18]]</td><br />
<td>[[Alberta/Calender/July#July_19|19]]</td><br />
<td>[[Alberta/Calender/July#July_20|20]]</td><br />
<td>[[Alberta/Calender/July#July_21|21]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_22|22]]</td><br />
<td>[[Alberta/Calender/July#July_23|23]]</td><br />
<td>[[Alberta/Calender/July#July_24|24]]</td><br />
<td>[[Alberta/Calender/July#July_25|25]]</td><br />
<td>[[Alberta/Calender/July#July_26|26]]</td><br />
<td>[[Alberta/Calender/July#July_27|27]]</td><br />
<td>[[Alberta/Calender/July#July_28|28]]</td><br />
</tr><br />
<tr><br />
<td>[[Alberta/Calender/July#July_29|29]]</td><br />
<td>[[Alberta/Calender/July#July_30|30]]</td><br />
<td>[[Alberta/Calender/July#July_31|31]]</td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<td></td><br />
<br />
<br />
</tr><br />
</table></td><td><br />
</td></tr></table></div><br />
<br />
<br />
<br />
To [[Alberta/Calender/August|August 2007]]<br><br />
Back to [[Alberta|UofA iGEM Home]]<br><br />
<br />
== July 1 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 2 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 3 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 4 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.<br />
<br />
-ED, JG, ML, MC, VH, AL, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 5 ==<br />
<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 6 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformation of E1010 did not work, it is in a Kan vector. SMRT.<br />
<br />
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.<br />
<br />
-ED<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 7 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.<br />
<br />
-JP,JB,AL<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 8 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.<br />
<br />
Pour Kan and Amp plates. Make TBE and TAE buffer.<br />
<br />
-ED, JP, MC, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 9 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.<br />
<br />
-ED, VH, JG<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 10 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.<br />
<br />
-ED, MC, AF, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 11 ==<br />
<br />
'''Lab Notes:'''<br />
<br />
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.<br />
<br />
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.<br />
<br />
<center> [[image: Alberta_e1010xlgold_july11.jpg]] <br />
<br />
E1010 XLGOLD 10 July 11</center><br />
<br />
-ED, MC, NG, JG, VH, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 12 ==<br />
<br />
'''General Notes:'''<br />
<br />
Calender up and running.<br />
<br />
Meeting tonight at 7:00 in CSC 2-49.<br />
<br />
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!!<br />
-Justin<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.<br />
<br />
Streak for single colonies of E1010, A0500, J61003.<br />
<br />
Purification of gel slice of digested J61003.<br />
<br />
<br />
<center> [[image: Alberta_e1010DH5a_july12.jpg]] <br />
<br />
E1010 DH5a July 12</center><br />
<br />
-ED, JG, AL, JB, MC, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 13 ==<br />
<br />
'''General Notes:'''<br />
<br />
Everyone who is coming tonight please be present 7:00pm sharp<br />
<br />
'''Lab Notes:'''<br />
<br />
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.<br />
<br />
-ED, JB<br />
<br />
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"<br><br />
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)<br><br />
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL<br><br />
Made some Kan plates<br><br />
<br />
-MC, JP, JG, CZ<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 14 ==<br />
<b>Schedule:</b><br />
<br />
AL,JP,NK<br />
<br />
<b>General Notes:</b><br />
<br />
If you are scheduled during the day please be in the lab at 1230hrs (PM) <br />
Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911<br />
<br />
Things to be continued today (CZ)<br><br />
1. Put LB plates in 4 degree fridge <br><br />
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control<br><br />
3. Xba1/EcoRI double digest <br><br />
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour. <br><br />
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.<br><br />
<br />
If you need anything else call me(Celine). Number on the blackboard. Thank you.�<br />
<br />
<b/>Lab Notes:</b><br />
JP, AL, NK, JB(?) <br><br />
<br />
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20 <br><br />
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20 <br><br />
3. Placed Kan plates at +4 <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 15 ==<br />
<b>Schedule:</b><br />
<br />
Jp,JB,AL<br />
<br />
<b>General Notes:</b><br />
<br />
Options: <br><br />
<br />
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)<br><br />
2. gel purify J61003 restriction products (promoter restriction sample) <br><br />
3. fundraising letters! <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow<br><br />
2. glycerol stocks of Chlorobium tepidum<br><br />
3. finished grant application forms<br><br />
- JP, AL, NG, MC<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 16 ==<br />
<b>Schedule:</b><br />
<br />
CZ,VH,AF<br />
<br />
<b>General Notes:</b><br />
<br />
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP) <br><br />
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining <br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed gel extraction of E,X digested J61003. (in box in -20)<br />
<br />
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.<br />
<br />
-VH, ED, CZ<br />
<br />
<br />
'''Quotes of the Day:'''<br />
<br />
"If I had a remote, I'd change the cd."<br />
<br />
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it<br />
<br />
"I would never drive this tired. But I WILL operate chemicals."<br />
<br />
Anonymous<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 17 ==<br />
<b>Schedule:</b><br />
<br />
ED,VH,AF<br />
<br />
'''Lab Notes:'''<br />
<br />
Gel extracted E1010 from yesterday's slices.<br />
<br />
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post. <br><br />
<b> It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC </b> <br><br />
<br />
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED<br />
<br />
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.<br />
<br />
-ED,JG,VH,AF<br />
<br />
'''Quote of the Day'''<br />
<br />
"Cancer is an urban myth."<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 18 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,MC,NG<br />
<br />
<b>Lab Notes:</b><br />
<br />
Transformed Ligations 1,2,3,4 into XL10 Gold Competent cells <br><br />
Plated 1ul,10 ul, and 200ul on AMP <br><br />
Incubated @ 37celsius overnight<br><br />
<br />
-MC, NG, CZ<br />
<br />
<b>Quote(s) of the Day:</b><br />
<br />
"Asuuuuuuuuuuuum!!" <br><br />
In response to the awesome; "You're wayyyy to exciting for me right now" <br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 19 ==<br />
<br />
Meeting at 7:00, CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
ED,MC,JG<br />
<br />
<b>General Notes</b><br />
<br />
New Schedule is up<br><br />
check out the jar o' love @ the front :) <br><br />
<br />
<b>Lab notes</b><br />
<br />
Started 8X5mL overnights to test colonies of the RFP ligation. Hopefully JB will be available to do minipreps tomorrow. Miniprep protocol is tucked in Master Book.<br />
<br />
-ED,JG,MC<br />
<br />
<b>Quote of the Day </b><br />
<br />
"There is a pee throw up smell it the washroom"<br />
<br />
Reply<br />
<br />
"Someone must be pregnant"<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 20 ==<br />
<b>Schedule:</b><br />
<br />
JG,ED,MC <br><br />
<br />
<b>General Notes:</b><br><br />
<br />
There is now a Jar O' Love in the lab; Please do not drink chemicals from said jar o'love. Check it out - it's at the front on the instructors bench. It's to give shout outs for people who are rocking out and 'taking one for the team'. <br><br />
<br />
<b>Lab Notes:</b><br />
<br />
Miniprep Lig3 (8 samples) overnights - JB<br><br />
Made 1L LB broth (16 bottles) at the back to be put away in the sterile cupboards tomorrow <br><br />
Ran Gel of JB's Minipreps after digestion with Xba and Spe. All of the colonies were positive (band at 1.2 kB).<br><br />
Did some dishes - we should learn how to use the dishwasher<br><br />
<br />
- MC, ED, JG <br><br />
<br />
<b>Quote(s) of the day:</b><br><br />
<br />
"Always use protection when using the Jar O' Love"<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 21 ==<br />
<b>Schedule:</b><br />
<br />
JP,NK,AL<br />
<br />
<b>General Notes:</b><br />
<br />
<br />
Since it doesn't look like Nik will be showing up, maybe just call Alex and see if it works for him.<br />
<br />
All of the ligations of RFP into J61003 turned out. They are labelled JB July 19 1-8 in the DNA box. Transform 1 of them into XL10 Gold and plate on Amp with Tetracycline plates. You will have to add the Tet to the plates beforehand. I don't know what concentraion, but James suggested 1 ug/mL. Try plating different amounts.<br />
<br />
The reason that I am making these suggestions is that I read that XL10Golds are Tet resistant, so we can maybe induce the expression of RFP with the Tet because it is under the Tet promotor and then we can see the RFP.<br />
<br />
As always, call me if you need anything.<br />
<br />
-ED<br />
<br />
<b>Lab Notes:</b><br />
<br />
1. got into building <br><br />
2. waited for peers - no shows <br><br />
3. tried to break into room by climbing through roof (cement goes all the way up) <br><br />
4. went home... will complete work tomorrow - sorry i'll get swipe access asap... <br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 22 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,AL,NG<br />
<br />
<b>General Notes:</b><br />
<br />
How does 7:00pm sound?<br />
<br />
<b>Laboratory Notes:</b><br />
<br />
Transformed XL10 gold cells with J61003 (with RFP ligated in). Split cell aliquot into two 50microL aliquots and added 1microL DNA from ligation sample 1 and from ligation sample 4. Covered AMP plates with 50microL 1microg/ml tet, spread, let sit. Plated (1) 50microL sample and (1) 5microL + 45microL LB onto Tet/Amp plates for #'s 1&4. They are in 37C incubator. The remaining transformed cells are in +4C labeled July 22. <br><br />
<br />
<b>Serious Quote of the Month</b><br />
<br />
<b>Faust</b><br><br />
Then shall I see, with vision clear, <br><br />
How secret elements cohere, <br><br />
And what the universe engirds, <br><br />
And give up huckstering with words <br><br />
<br />
-Johann Wolfgang von Goethe <br><br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 23 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,CZ,JG<br />
<br />
<b>Laboratory Notes:</b><br><br />
1. Add more Tet on ligation plates from July 22. Hopefully red colonies will appear!<br><br />
2. Transformation of XL10Gold with<br><br />
- Butanol CoA dehydrogenase (1 in 1000 dilution)<br><br />
- Butenoyl CoA dehydrogenase (1 in 1000 dilution)<br><br />
- J23018 (RFP positive control)<br><br />
<br />
20 and 100 microlitres of transformed culture plated on Amp plate and incubate at 37 degrees<br><br />
<br />
The leftover mixture is stored in 4 degree fridge in case of no growth.<br><br />
3. Repeate overnight ligation from July 22. The exact amounts of insert, vector etc. are on the blackboard.<br><br />
<br />
For tomorrow:<br><br />
1. Pick colonies for transformed plates.<br><br />
2. Plate ligation mixtures with appropriate amount of Tet.<br><br />
3. Observe red colonies from the July 22 ligation plates if the cells are still alive.<br><br />
<br />
Memorable Quote:<br><br />
Don't force it. Let it come out naturally.<br><br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 24 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,NG<br />
<br />
'''Lab Notes:'''<br />
<br />
Transformed I0500 (arabinose promotor) into HB101 competent cells. Plated 2, 20, 200 uL on Kan.<br />
<br />
Transformed ligations from yesterday into XL10 gold. Plated 100 uL on Amp.<br />
<br />
RFP control from yesterday worked, but sadly our ligations are not expressing any RFP. It was still exciting to see red control cells. It seems as if the Tet promotor is induced at a low level without any Tet.<br />
<br />
Started 4X5ml overnights of enoyl coA hydratase and butyryl coa DH. There seems to be some confusion with the naming of these proteins. These are the names that I have used in the BioBricks. We will be able to tell if they are correctly labelled when we do the digests. Enoyl coa hydratase is about 840 bp and butyryl coa DH is about 1200 bp.<br />
<br />
Note on agarose gels: Use only the wide combs (8 lanes) because we are having to many problems with the narrow lanes. This may mean that you will need to pour multiple gels (oh my). Also, try and run two ladders per gel until we get the ladder problem sorted out. Also, make sure that you zoom in on the gel when taking a picture.<br />
<br />
Note on Tet: should be stored 12.5 mg/mL in 50% ethanol, 50% water solution. This is a 1000X solution. Keep this in mind when using Tet again.<br />
<br />
-ED, AF, James<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 25 ==<br />
<br />
<b>Schedule:</b><br />
<br />
Jp,MC,VH<br />
<br />
<b>General Notes:</b><br />
<br />
Use of wide combs- 5microL of sample works well with 3.5microL of ladder. Make sure you stir up the ladder before use!<br><br />
<br />
'''Lab Notes:'''<br />
<br />
Completed minipreps of enoylcoa hydratase and butyryl coa dh. They are in the new box with the green tape. I think that there are two types of digests we should be doing: one to make sure we have the right stuff (digest with EcoR1 and Pst1) and one to clone in the RBS. To clone in the RBS, cut B0034 with Eco and Spe and cut the enoyl coa hydratase and butyryl coa dh with Eco and Xba.<br />
<br />
Check to see if any colonies have grown on I0500 plates in incubator. There weren't any visible on Adam's transformations this morning. James also transformed some this morning, so they may be hard to see. If there are colonies, start overnights and see if Jori can do minipreps tomorrow. If Jori can't make it, get ahold of me and I can. AND if there are no colonies, just post here and I'll go check the plates in the morning.<br />
<br />
I also forgot to start glycerol stocks of our enoly coa hydratase and butyryl coa dh. If you guys could restart some overnights from the plates in the fridge so I can do it tomorrow that would be great.<br />
<br />
I had to rush to work this morning and didn't get any of this written in the lab book. So you guys can make fun of me or whatever. I guess I belong in the Science Hall of Shame.<br />
<br />
-ED <br><br />
<br />
Enny = Enoly CoA Hydratase<br><br />
Benny = Butytyl CoA DH<br />
<br />
Checked plates<br><br />
- Adam - no growth: parafilmed and put in 4 fridge<br><br />
- James'? J23018: red colonies parafilmed and put in 4 fridge (was this the control?)<br><br />
<br />
Overnights of Benny & Enny<br><br />
- 3x 5mlLB each <br><br />
<br />
Restrictions<br><br />
- B0034 with SPE & ECO<br><br />
- Enny with Xba & ECO<br><br />
-Benny with Xba & ECO<br><br />
These are stored in the -20 freezer<br><br />
<br />
- MC, NK, VH, NG, JP<br><br />
<br />
<b>For Tomorrow:</b><br><br />
- Glycerol stocks of Enny & Benny (these colonies are likely different from the transformations as it was not marked out which colonies were picked for transformations)<br><br />
- Gels for restrictions of B0034, Enny & Benny<br />
- cut out the bands; gel extractions? (if there is time)<br><br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 26 ==<br />
<br />
Meeting at 7:00, Room CAB 373.<br />
<br />
<b>Schedule:</b><br />
<br />
AL,VH,ED<br />
<br />
<b>Lab Book</b><br />
<br />
Ran gel of "Enny", "Benny" and Boo IV against 1 kb ladder.<br />
<br />
Cleared up glasswares in the autoclave.<br />
<br />
Parafilm all unparafilmed plates.<br />
<br />
Checked out red fluorescence of J23018.<br />
<br />
Attempted working on fundraising letters (?)<br />
<br />
-ED, VH, AL, JG, MC, AF, Doug<br />
<br />
<b>Favourite "F" word of the day:</b><br />
<br />
fulcrum<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 27 ==<br />
<br />
<b>Schedule:</b><br />
<br />
JP,MC,NK<br><br />
The theme is Foul music Friday - bring any bad music you have (or embarassed that you have in your collection)<br />
<br />
<b>General Notes:</b><br />
<br />
1. This is whats on the agenda for this evening:<br />
[[Image: Alberta_promoterprob.jpg]]<br />
<br />
<b>Lab book:</b><br />
<br />
Digest Enny(1,4), Benny (1,2) B0034(2,4) with PST, XBA, SPE<br><br />
Enny with PST and XBA<br><br />
Benny with PST and XBA<br><br />
B0034 with PST and SPE<br><br />
O/N of Benny and Enny glycerol stock (3 each)<br><br />
<br />
'''''Note the 4 degree fridge at the back is out of commision. All iGEM materials have been moved to the room where we take pictures of gels. The door labelled IGEM.''''' Nik is here on saturday and Celine can show on sunday.<br><br />
<br />
- JG, NK, MC, JP, and James (cos he loves us) <br />
<br />
<br />
<b>Jame's Fun Tip of the Day:</b><br />
If you want to run a gel for a shorter period of time ad the EtBR to the gel rather than the buffer. You can add ~4ul to the gel rather than 10ul to the buffer. <br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 28 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,JG,NK<br />
<br />
Are we meeting at noon? Please call me before noon to confirm. Thank you. -Celine<br />
<br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Glycerol stocks of Enny & Benny from the O/N<br><br />
- Gels of Enny, Benny & B0034 (put the EtBr in the gel)<br><br />
- Ligations of Enny+Benny into B0034 with T4 ligase; leave O/N in incubator at the back room (near the phone) set at 13degrees celsius<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 29 ==<br />
<br />
<b>Schedule:</b><br />
<br />
CZ,AF,NG<br><br />
<br />
<b>General Notes:</b><br />
<br />
To Do:<br><br />
- Transformation of Ligations into XL10 Gold<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 30 ==<br />
<br />
<b>Schedule:</b><br />
<br />
VH,JP,JG,<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
== July 31 ==<br />
<br />
<b>Schedule:</b><br />
<br />
AF,ED,MC<br />
<br />
<br />
[[Alberta/Calender/July#July|to the top]]<br />
<br />
<br />
<br />
[[Alberta|UofA iGEM Home]]<br><br />
[[Alberta/Calender/June|To June 2007]]<br><br />
[[Alberta/Calender/August|To August 2007]]</div>CelineYZ