2/6/2012 To-Do
Summary of Weekend Progress
- Restriction map confirmed that the restriction enzymes E, X, S all work
- 4% gel confirmed the annealing of the ultramers for J85010, J85012, and J85013
- Gel of the EX digests of J85201 and J06702; and the ES digests of S03119 and Chlor confirmed expected band lengths.
- Gel extraction of the parts yielded good DNA concentrations, with good 260/230 ratio
- Phosphatase->Gel extract->Ligate->Tranformed
- Transformation results, however, yielded no colonies for all the negative controls and ligations. Postitive controls yielded conolonies
- Because the ultramer annealing was confirmed, the ligation step must be the problem.
- Will retry ligation following advice: http://openwetware.org/wiki/DNA_ligation
Today's Workflow
- Miniprep J06702
- PCR J06702 and 069 fragment with recently ordered Gibson primers J85014, J85015
- Gibson assemble the PCR products
- Reattempt ligation
- Inoculate additional S03119.
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