Imperial/Cell by Date/Implementation

From 2007.igem.org

< Imperial(Difference between revisions)
(DNA Constructs)
m
 
(15 intermediate revisions not shown)
Line 1: Line 1:
{{Template:IC07navmenu}}
{{Template:IC07navmenu}}
-
__NOTOC__
+
<br clear="all">
 +
<html>
 +
<link rel="stylesheet" href="/igem07/index.php?title=User:Dirkvs/Stylesheets/IC07persist.css&action=raw&ctype=text/css" type="text/css" />
-
== DNA Constructs ==
+
<div id="tabs">
-
 
+
  <ul>
-
{|border=1 width=90% align="center"
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Introduction" title=""><span>Introduction</span></a></li>
-
|-
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Specification" title=""><span>Specifications</span></a></li>
-
|width="30%"|<center>[[Image:Imperial CBD Design CBD1.png ]]</center>
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Design" title=""><span>Design</span></a></li>
-
|width="--"|Notes: This part can be found as [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] in the registry
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Modelling" title=""><span>Modelling</span></a></li>
-
|-
+
    <li><a class="current" href="https://2007.igem.org/Imperial/Cell_by_Date/Implementation" title=""><span>Implementation</span></a></li>
-
|<center>[[Image:Imperial CBD Design CBD2.png]]</center>
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Testing" title=""><span>Testing</span></a></li>
-
|width="--"|Notes: the part [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] we found in the registry was not functional, therefore, we made our own T7 promoter and ligated it to the GFP expression biobrick [http://partsregistry.org/Part:BBa_I13504 BBa_I13504]
+
    <li><a href="https://2007.igem.org/Imperial/Cell_by_Date/Conclusion" title=""><span>Conclusion</span></a></li>
-
|}
+
  </ul>
 +
</div>
 +
<hr />
<br clear="all">
<br clear="all">
 +
</html>
 +
__NOTOC__
 +
= Cell by Date: Implementation =
-
===Constitutive tetR Promoter===
 
-
<center>[[Image:Imperial CBD Design CBD1.png]]</center>
 
-
'''Ligation Strategy:'''
 
-
N/A
+
{|border=1 width=100% align="center" cellpadding="10px"
-
 
+
-
'''Quality Control:'''
+
-
{|border=1 width=90% align="center"
+
|-
|-
-
|width="20%"|&nbsp;Testing ''in vivo''
+
|colspan="3" style="background-color:#eeeeff"| <center>'''DNA Construct Implementation'''</center>
-
|width="10%" align="center"|Done
+
|- align="left"
-
|width="--"|Results can be found at -----
+
|wifth="20%" style="background-color:#ddeeff" align="center"| '''Construct Design'''
 +
|width="40%"| '''Concept 1'''<br />[[Image:IC07_design_CBDcon1.png|center|370px]]
 +
|width="40%"| '''Concept 2'''<br />[[Image:IC07_design_CBDcon2.png|center|370px]]
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 1: Availability''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"| Found as [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] in Registry
 +
|width="40%"| Found as [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] in Registry.
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 2: Cloning / Amplification''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"| [[Image:IC07_ID2-cloning.png|thumb|center|290px| BBa_I13522 - Lane R2 (937bp + 2079bp vector)]]
 +
|width="40%"| [[Image:IC07_CBD1-cloning.png|thumb|center|290px| BBa_E7104 - Lane 11 (Missing 826bp)]]
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 3: Ligation''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"| Not Required
 +
|width="40%"| Since [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] was not functional, we made our own promoter and and ligated it to [http://partsregistry.org/Part:BBa_I13504 BBa_I13504]
 +
<br>
 +
{| width="80%"
|-
|-
-
|&nbsp;Testing ''in vitro''
+
|[[Image:Imperial_CBD_Design_CBD4.png|37px]]
-
|align="center"|Done
+
-
|Results can be found at the [[Imperial/Cell_by_Date/Testing|testing]] page.
+
-
|-
+
-
 
+
-
|&nbsp;Sequencing
+
-
|align="center"|Pending
+
-
|''Sequence results not obtained yet''
+
-
|}
+
-
 
+
-
===Constitutive T7 Promoter===
+
-
<center>[[Image:Imperial CBD Design CBD2.png]]</center>
+
-
 
+
-
'''Ligation Strategy:'''
+
-
{|
+
-
|-
+
-
|[[Image:Imperial_CBD_Design_CBD4.png]]
+
|<font size="+3">+</font>
|<font size="+3">+</font>
-
|[[Image:Imperial_CBD_Design_CBD3.png]]
+
|[[Image:Imperial_CBD_Design_CBD3.png|110px]]
|<font size="+3">=</font>
|<font size="+3">=</font>
-
|[[Image:Imperial CBD Design CBD2.png]]
+
|[[Image:Imperial CBD Design CBD2.2.png|130px]]
|}
|}
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 4: Purification of Activator Proteins''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"| Not Required
 +
|width="40%"| Not Required
 +
|-
 +
|width="20%" style="background-color:#ddeeff" align="center" | '''Stage 5: Quality Control''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"| [[Image:IC2007_Experiments_Phase1_Protocol1-1.jpg|center|350px]]<br />
 +
* '''Testing ''in vivo''''': Passed ([[Imperial/Wet_Lab/Protocols/CBD1.1|Protocols]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/CBD1.1 Results])
 +
* '''Sequencing of Construct''': Completed
 +
|width="40%"| [[Image:IC_2007_PT7-invitro.png|center|350px]]<br />
 +
* '''Testing ''in vitro''''': Passed ([[Imperial/Wet_Lab/Protocols/Prot1.9|Protocols]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/Res1.9 Results])
 +
* '''Sequencing of Construct''': Completed
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 6: Final Implementation Status'''
 +
|width="40%" style="background-color:#b8efb0"| Ready for Testing
 +
|width="40%" style="background-color:#b8efb0"| Ready for Testing
 +
|}
 +
<br clear="all">
 +
-
'''Quality Control:'''
+
{|border=1 width=100% align="center" cellpadding="10px"
-
{|border=1 width=90% align="center"
+
|-
|-
-
|width="20%"|&nbsp;Testing ''in vivo''
+
|colspan="3" style="background-color:#eeeeff"| <center>'''Chassis Construct Implementation'''</center>
-
|width="10%" align="center"|Done
+
|- align="center"
-
|width="--"|Results can be found at -----
+
|width="20%" style="background-color:#ddeeff"| '''Chassis Design'''
-
|-
+
|colspan="2" align="center" |[[Image:IC07_CFS_components.png|300px]]
-
|&nbsp;Testing ''in vitro''
+
|- align="center"
-
|align="center"|Done
+
|width="20%" style="background-color:#ddeeff"| '''Stage 1: Availability''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
-
|Results can be found at -----
+
|width="40%"| Possibility of making Homemade S30 Cell Extract.
-
|-
+
|width="40%"| Commercially available S30 Cell Extract purchasable from [http://www.promega.com/search/Default.aspx?query=s30+extract Promega].
-
 
+
|- align="center"
-
|&nbsp;Sequencing
+
|width="20%" style="background-color:#ddeeff"| '''Stage 2: Preparation''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
-
|align="center"|Pending
+
|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.1|Protocol]])
-
|''Sequence results not obtained yet''
+
|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.3|Protocol]])
 +
|-  
 +
|width="20%" style="background-color:#ddeeff" align="center" | '''Stage 3: Quality Control''' <br /> <br> [[Image:Icegems-Devcycle-downarrow.png|30px]]
 +
|width="40%"|
 +
* '''Testing ''in vitro''''': Failed ([[Imperial/Wet_Lab/Protocols/CE1.5|Protocol]] and [[Imperial/Wet_Lab/Results/Chas1.1 |Results]])
 +
|width="40%"|
 +
* '''Testing ''in vitro''''': Passed ([[Imperial/Wet_Lab/Protocols/CBD1.2|Protocol]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/CBD1.2 Results])
 +
|- align="center"
 +
|width="20%" style="background-color:#ddeeff"| '''Stage 4: Final Implementation Status'''
 +
|width="40%" style="background-color:#ddb99e"| Aborted. <br /> Expression capacity of chassis was low.
 +
|width="40%" style="background-color:#b8efb0"| Ready for Testing
|}
|}
<br clear="all">
<br clear="all">
-
== Chassis ==
 
-
cell extract preparation protocols.
+
 
 +
<center> [https://2007.igem.org/Imperial/Cell_by_Date/Modelling << Modelling ] | Implementation | [https://2007.igem.org/Imperial/Cell_by_Date/Testing Testing/Validation >>]</center>

Latest revision as of 02:35, 27 October 2007




Cell by Date: Implementation

DNA Construct Implementation
Construct Design Concept 1
IC07 design CBDcon1.png
Concept 2
IC07 design CBDcon2.png
Stage 1: Availability

Icegems-Devcycle-downarrow.png
Found as [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] in Registry Found as [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] in Registry.
Stage 2: Cloning / Amplification

Icegems-Devcycle-downarrow.png
BBa_I13522 - Lane R2 (937bp + 2079bp vector)
BBa_E7104 - Lane 11 (Missing 826bp)
Stage 3: Ligation

Icegems-Devcycle-downarrow.png
Not Required Since [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] was not functional, we made our own promoter and and ligated it to [http://partsregistry.org/Part:BBa_I13504 BBa_I13504]


Imperial CBD Design CBD4.png + Imperial CBD Design CBD3.png = Imperial CBD Design CBD2.2.png
Stage 4: Purification of Activator Proteins

Icegems-Devcycle-downarrow.png
Not Required Not Required
Stage 5: Quality Control

Icegems-Devcycle-downarrow.png
IC2007 Experiments Phase1 Protocol1-1.jpg

  • Testing in vivo: Passed (Protocols and Results)
  • Sequencing of Construct: Completed
IC 2007 PT7-invitro.png

  • Testing in vitro: Passed (Protocols and Results)
  • Sequencing of Construct: Completed
Stage 6: Final Implementation Status Ready for Testing Ready for Testing



Chassis Construct Implementation
Chassis Design IC07 CFS components.png
Stage 1: Availability

Icegems-Devcycle-downarrow.png
Possibility of making Homemade S30 Cell Extract. Commercially available S30 Cell Extract purchasable from [http://www.promega.com/search/Default.aspx?query=s30+extract Promega].
Stage 2: Preparation

Icegems-Devcycle-downarrow.png
Preparation: Done (Protocol) Preparation: Done (Protocol)
Stage 3: Quality Control

Icegems-Devcycle-downarrow.png
Stage 4: Final Implementation Status Aborted.
Expression capacity of chassis was low.
Ready for Testing



<< Modelling | Implementation | Testing/Validation >>