IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/28: Difference between revisions
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- run a gel | - run a gel | ||
[[Image:photoq.gif| | [[Image:photoq.gif|150px|center]] | ||
* Result : nothing on the gel!!! But we can not manage to PCR anything... there is a problem in our PCR protocol, or products! | * Result : nothing on the gel!!! But we can not manage to PCR anything... there is a problem in our PCR protocol, or products! | ||
Revision as of 06:30, 5 September 2008
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Results from transformation of ligation products- nothing on plate, even on control plate
- cells non competent anymore (try with fresh competent cells) - not enough DNA (try with 10μL of DNA) - very short ligation products Results from Bacillus transformation with glycerol stocks
Transformation of ligation products (new)
- new fresh competent TOP 10 - 5μL of DNA (1.5μL of PUC9) - 2h30 in the incubator Test AmyE insertion
- 20μL of lysozyme, a few colonies for each - Cycle : 15min qt 37°C, 15min at 99°C, 1min at 4°C, 1min at 99°C and 1min at 4°C - for each PCR : 12μL of cells, 1μL of cells, 1μL of primer1, 1μL of primer2, 5μL of master mix - PCR (iGEM program) - run a gel 
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