Imperial/Cell by Date/Implementation

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