Imperial/Cell by Date/Implementation

From 2007.igem.org

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|colspan="3" style="background-color:#eeeeff"| <center>'''DNA Construct Implementation'''</center>
|colspan="3" style="background-color:#eeeeff"| <center>'''DNA Construct Implementation'''</center>
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|wifth="20%" style="background-color:#ddeeff"| '''Construct Design'''
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|wifth="20%" style="background-color:#ddeeff" align="center"| '''Construct Design'''
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|width="40%"| [[Image:Imperial CBD Design CBD1.png]]
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|width="40%"| '''Concept 1'''<br />[[Image:Imperial CBD Design CBD1.png|center]]
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|width="40%"| [[Image:Imperial CBD Design CBD2.png]]
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|width="40%"| '''Concept 2'''<br />[[Image:Imperial CBD Design CBD2.png|center]]
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|width="20%" style="background-color:#ddeeff"| '''Stage 1: Availability'''
|width="20%" style="background-color:#ddeeff"| '''Stage 1: Availability'''
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|width="20%" style="background-color:#ddeeff" align="center" | '''Stage 5: Quality Control'''
|width="20%" style="background-color:#ddeeff" align="center" | '''Stage 5: Quality Control'''
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* '''Testing ''in vivo''''': Done ([[Imperial/Wet_Lab/Protocols/ID1.1|Protocol]] and [http://openwetware.org/index.php?title=IGEM:Imperial/2007/Wet_Lab/results/ID1.1 Results])
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* '''Testing ''in vivo''''': Done ([[Imperial/Wet_Lab/Protocols/CBD1.1|Protocols]] and [http://openwetware.org/index.php?title=IGEM:Imperial/2007/Wet_Lab/results/CBD1.1 Results])
* '''Sequencing of Construct''': Pending
* '''Sequencing of Construct''': Pending
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|width="40%"|  
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* '''Testing ''in vivo''''': Done ([[Imperial/Wet_Lab/Protocols/Prot1.7|Protocol]] and [http://openwetware.org/wiki/IGEM:Imperial/2007/Wet_Lab/results/Res1.7 Results])  
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* '''Testing ''in vivo''''': Done ([[Imperial/Wet_Lab/Protocols/CBD1.1|Protocols]] and [http://openwetware.org/index.php?title=IGEM:Imperial/2007/Wet_Lab/results/CBD1.1 Results])
* '''Sequencing of Construct''': Pending
* '''Sequencing of Construct''': Pending
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== DNA Construct Status ==
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{|border=1 width=90% align="center"
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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>'''Chassis 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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|width="20%" style="background-color:#ddeeff"| '''Chassis Design'''
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|<center>[[Image:Imperial CBD Design CBD2.png]]</center>
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|colspan="2" align="center" |[[Image:IC07_CFS_components.png|300px]]
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|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]
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|width="20%" style="background-color:#ddeeff"| '''Stage 1: Availability'''
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|width="40%"| Possibility of making Homemade S30 Cell Extract.
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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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|width="20%" style="background-color:#ddeeff"| '''Stage 2: Preparation'''
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|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.1|Protocol]])
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|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.3|Protocol]])
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|width="20%" style="background-color:#ddeeff" align="center" | '''Stage 3: Quality Control'''
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* '''Testing ''in vitro''''': Done (Protocol and Results)
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* '''Testing ''in vitro''''': Done ([[Imperial/Wet_Lab/Protocols/CBD1.2|Protocols]] and [http://openwetware.org/index.php?title=IGEM:Imperial/2007/Wet_Lab/results/CBD1.2 Results])
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|width="20%" style="background-color:#ddeeff"| '''Stage 6: Final Implementation Status'''
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|width="40%" style="background-color:#ddb99e"| Aborted. <br /> Expression capacity of chassis was low.
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|width="40%" style="background-color:#b8efb0"| Ready for Testing
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===Constitutive tetR Promoter===
 
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<center>[[Image:Imperial CBD Design CBD1.png]]</center>
 
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'''Ligation Strategy:'''
 
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N/A
 
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'''Quality Control:'''
 
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|width="20%"|&nbsp;Testing ''in vivo''
 
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|width="10%" align="center"|Done
 
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|width="--"|Results can be found at -----
 
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|&nbsp;Testing ''in vitro''
 
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|align="center"|Done
 
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|Results can be found at the [[Imperial/Cell_by_Date/Testing|testing]] page.
 
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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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===Constitutive T7 Promoter===
 
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<center>[[Image:Imperial CBD Design CBD2.png]]</center>
 
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'''Ligation Strategy:'''
 
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|[[Image:Imperial_CBD_Design_CBD4.png]]
 
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|[[Image:Imperial_CBD_Design_CBD3.png]]
 
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|<font size="+3">=</font>
 
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|[[Image:Imperial CBD Design CBD2.png]]
 
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'''Quality Control:'''
 
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{|border=1 width=90% align="center"
 
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|width="20%"|&nbsp;Testing ''in vivo''
 
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|width="10%" align="center"|Done
 
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|width="--"|Results can be found at -----
 
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|&nbsp;Testing ''in vitro''
 
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|align="center"|Done
 
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|Results can be found at -----
 
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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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== Chassis Status ==
 
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cell extract preparation protocols.
 
== Navigation==
== Navigation==
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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>
<center> [https://2007.igem.org/Imperial/Cell_by_Date/Modelling << Modelling ] | Implementation | [https://2007.igem.org/Imperial/Cell_by_Date/Testing Testing/Validation >>]</center>

Revision as of 13:35, 24 October 2007


Cell by Date: Implementation

DNA Construct Implementation
Construct Design Concept 1
Imperial CBD Design CBD1.png
 Concept 2
Imperial CBD Design CBD2.png
Stage 1: Availability Found as BBa_I13522 in Registry Found as BBa_E7104 in Registry.
Stage 2: Cloning / Amplification
BBa_I13522 - Lane R2 (937bp + 2079bp vector)
BBa_E7104 - Lane 11 (Missing 826bp)
Stage 3: Ligation Not Required Since BBa_E7104 was not functional, we made our own promoter and and ligated it to BBa_I13504


Imperial CBD Design CBD4.png + Imperial CBD Design CBD3.png = Imperial CBD Design CBD2.png
Stage 4: Purification of Activator Proteins Not Required Not Required
Stage 5: Quality Control
  • Testing in vivo: Done (Protocols and Results)
  • Sequencing of Construct: Pending
  • Testing in vivo: Done (Protocols and Results)
  • Sequencing of Construct: Pending
Stage 6: Final Implementation Status Ready for Testing Ready for Testing



Chassis Construct Implementation
Chassis Design IC07 CFS components.png
Stage 1: Availability Possibility of making Homemade S30 Cell Extract. Commercially available S30 Cell Extract purchasable from Promega.
Stage 2: Preparation Preparation: Done (Protocol) Preparation: Done (Protocol)
Stage 3: Quality Control
  • Testing in vitro: Done (Protocol and Results)
Stage 6: Final Implementation Status Aborted.
Expression capacity of chassis was low. 		Ready for Testing



Navigation

<< Modelling | Implementation | Testing/Validation >>
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