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Team:Brown/Project/Light pattern/Workflow
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==Workflow== | ==Workflow== | ||
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| + | Flesh this out! | ||
| + | Starts with design of circuit, describes how modeling was started and why. Describes obtaining of DNA material from all over the place. 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, 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. | ||
Revision as of 11:40, 27 October 2010
Light-Pattern Controlled Circuit
Workflow
Flesh this out! Starts with design of circuit, describes how modeling was started and why. Describes obtaining of DNA material from all over the place. 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, 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.
