Imperial/Infector Detector/Implementation

From 2007.igem.org

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  <ul>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Introduction" title=""><span>Introduction</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Specification" title=""><span>Specifications</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Design" title=""><span>Design</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Modelling" title=""><span>Modelling</span></a></li>
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    <li><a class="current" href="https://2007.igem.org/Imperial/Infector_Detector/Implementation" title=""><span>Implementation</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Testing" title=""><span>Testing</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/F2620 Comparison" title=""><span>F2620</span></a></li>
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    <li><a href="https://2007.igem.org/Imperial/Infector_Detector/Conclusion" title=""><span>Conclusion</span></a></li>
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__NOTOC__
= Infector Detector: Implementation =
= Infector Detector: Implementation =
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== DNA Construct Status ==
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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="50%"|<center>[[Image:Imperial ID Design ID1.png]]</center>
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|colspan="3" style="background-color:#eeeeff"| <center>'''DNA Construct Implementation''' </center>
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|width="--"|&nbsp;Notes: This part can be found as [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] in the registry
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|- align="center"
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|wifth="20%" style="background-color:#ddeeff" border="0" | '''Construct Design'''
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|width="40%" align="left" | [[Image:IC07_design_ID.png|center|370px]]
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|width="40%" align="left" |  [[Image:IC07_design_ID-con2.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_T9002 BBa_T9002] in Registry. <br />[[Image:Imperial ID Design ID1.png|center|]]
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|width="40%"| Found as [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] in Registry.  <br /> [[Image:Imperial_ID_Design_ID2.png|center]]
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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_ID1-cloning.png|thumb|center|350px| BBa_T9002 - Lane 18 (1945bp)]]
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|width="40%"| [[Image:IC07_ID2-cloning.png|thumb|center|290px| BBa_J37032 - Lane R25 (938bp + 2079bp vector)]]
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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%"| Not Required
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|- align="center"
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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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|width="40%"| Not Required
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|width="40%"| [[Image:Purification_of_LuxR.PNG|center|thumb|200px|Purification of LuxR protein ([[Imperial/Wet_Lab/Protocols/Prot1.5|Protocol]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/Res1.5 Results])]]
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|-
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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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|width="40%"| [[Image:IC_2007_Plux_in_vivo.PNG|center|350px]]<br />
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* '''Testing ''in vivo''''': Passed ([[Imperial/Wet_Lab/Protocols/ID1.1|Protocol]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/ID1.1 Results])
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* '''Sequencing of Construct''': Partially Sequenced
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|width="40%"| [[Image:IC_2007_LuxR_testing_1.PNG|center|350px]]<br />
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* '''Testing ''in vitro''''': Failed ([[Imperial/Wet_Lab/Protocols/Prot1.7|Protocol]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/ID1.7 Results])
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* '''Sequencing of Construct''': Completed
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|- align="center"
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|width="20%" style="background-color:#ddeeff"| '''Stage 6: Final Implementation Status'''
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|width="40%" style="background-color:#b8efb0"| Ready for Testing
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|width="40%" style="background-color:#ddb99e"| Aborted. <br /> LuxR protein not functional.
|}
|}
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=== AHL Receiver + GFP Reporter ===
 
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<center>[[Image:Imperial ID Design ID1.png]]</center>
 
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'''Ligation Strategy:'''
 
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N/A
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{|border=1 width=100% align="center" cellpadding="10px"
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'''Quality Control:'''
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{|border=1 width=90% align="center"
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|-
|-
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|width="20%"|&nbsp;Testing ''in vivo''
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|colspan="3" style="background-color:#eeeeff"| <center>'''Chassis Construct Implementation'''</center>
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|width="10%" align="center"|Done
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|- align="center"
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|width="--"|Results can be found at -----
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|width="20%" style="background-color:#ddeeff"| '''Chassis Design'''
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|-
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|colspan="2" align="center" |[[Image:IC07_CFS_components.png|300px]]
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|&nbsp;Testing ''in vitro''
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|- align="center"
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|align="center"|Done
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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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|Results can be found at the [[Imperial/Infector_Detector/Testing|testing]] page.
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|width="40%"| Possibility of making Homemade S30 Cell Extract.
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|-
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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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|- align="center"
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|&nbsp;Sequencing
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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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|align="center"|Pending
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|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.1|Protocol]])
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|''Sequence results not obtained yet''
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|width="40%"| '''Preparation''': Done ([[Imperial/Wet_Lab/Protocols/CE1.3|Protocol]])
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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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|width="40%"|
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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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|width="40%"|
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* '''Testing ''in vitro''''': Passed ([[Imperial/Wet_Lab/Protocols/ID1.2|Protocol]] and [https://2007.igem.org/Imperial/Wet_Lab/Results/ID1.2 Results])
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|- align="center"
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|width="20%" style="background-color:#ddeeff"| '''Stage 4: 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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== Chassis Status ==
 
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cell extract preparation protocols
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<center>  [https://2007.igem.org/Imperial/Infector_Detector/Modelling << Modelling] | Implementation | [https://2007.igem.org/Imperial/Infector_Detector/Testing Testing/Validation >>]
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</center>

Latest revision as of 02:33, 27 October 2007



Infector Detector: Implementation

DNA Construct Implementation
Construct Design
IC07 design ID.png
IC07 design ID-con2.png
Stage 1: Availability

Icegems-Devcycle-downarrow.png 		Found as [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] in Registry.
Imperial ID Design ID1.png
 Found as [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] in Registry.
Imperial ID Design ID2.png
Stage 2: Cloning / Amplification

Icegems-Devcycle-downarrow.png
BBa_T9002 - Lane 18 (1945bp)
BBa_J37032 - Lane R25 (938bp + 2079bp vector)
Stage 3: Ligation

Icegems-Devcycle-downarrow.png 		Not Required Not Required
Stage 4: Purification of Activator Proteins

Icegems-Devcycle-downarrow.png 		Not Required
Purification of LuxR protein (Protocol and Results)
Stage 5: Quality Control

Icegems-Devcycle-downarrow.png
IC 2007 Plux in vivo.PNG

  • Testing in vivo: Passed (Protocol and Results)
  • Sequencing of Construct: Partially Sequenced
IC 2007 LuxR testing 1.PNG

  • Testing in vitro: Failed (Protocol and Results)
  • Sequencing of Construct: Completed
Stage 6: Final Implementation Status Ready for Testing Aborted.
LuxR protein not functional.



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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