"
Team:Brown/Project/Light pattern/Workflow
From 2010.igem.org
(→Circuit Beginnings) |
|||
| Line 6: | Line 6: | ||
===Circuit Beginnings=== | ===Circuit Beginnings=== | ||
| - | + | Once we had designed our circuit, we went about obtaining all of the necessary parts for its construction. Because the registry did not contain the majority of parts crucial to our circuit, we collaborated with teams EPF-Lausanne, PKU, and Davidson-Missouri to obtain the relevant parts. From EPF-Lausanne, we received <partinfo>BBa_K191003</partinfo> and <partinfo>BBa_K191005</partinfo>. We received from PKU Beijing... | |
| - | + | ||
| - | + | ||
| - | + | ||
===Experimental Procedure=== | ===Experimental Procedure=== | ||
describes how the expression construct for LOVTAP and the reporter for LOVTAP (WT LOVTAP R2) were transformed, double transformed to generally confirm function, | describes how the expression construct for LOVTAP and the reporter for LOVTAP (WT LOVTAP R2) were transformed, double transformed to generally confirm function, | ||
Revision as of 23:03, 27 October 2010
Light-Pattern Controlled Circuit
Workflow
Circuit Beginnings
Once we had designed our circuit, we went about obtaining all of the necessary parts for its construction. Because the registry did not contain the majority of parts crucial to our circuit, we collaborated with teams EPF-Lausanne, PKU, and Davidson-Missouri to obtain the relevant parts. From EPF-Lausanne, we received <partinfo>BBa_K191003</partinfo> and <partinfo>BBa_K191005</partinfo>. We received from PKU Beijing...
Experimental Procedure
describes how the expression construct for LOVTAP and the reporter for LOVTAP (WT LOVTAP R2) were transformed, double transformed to generally confirm function,
creation/ordering of primers for WT LOVTAP R2 to isolate the double repressor part and add bgl sites (since not compatible with RFC 10) and subsequent PCR (image of gel if we have) (Now named LOV2).
Describes unsuccessful transformations (3 times!) of Bistable part from PKU.
Transformation of CI, AraC, Mnt. Design and ordering primers to add bgl-prefix and RBS to the left, bgl-suffix to the right. PCR, image of gary's gel (this is in the lab),
ligation to pGEM and transformation successful. Colony PCR to test for insert inconclusive.
Decision that we should test Conversion module and memory module together with just CI to confirm function. Double digest and ligation of RFC 10 CI and Ter+Ter. Successful transformation, PCR on bgl prefix+RBS, bgl-suffix, successful PCR.
NOTE THAT THIS IS WHERE THE WE WERE AT AT THE WIKI FREEZE. NOTE THAT HERE IS WHAT WE HOPE TO HAVE BY THE COMPETITION: ligate PCR product with pGEM, transform, double digest bgl'd CI+TERTER and LOV2 in bgl standard, ligate, transform. Double digest and ligate LOVTAP expression upstream, transform. Transform bistable (with One Shot Invitrogen cells), double transform and test.
