User:Tk/Notebook/MF-xfm/2008/04/13

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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Transformation results

 * E. coli transforms at 400 colonies
 * No MF transformants on any plates
 * Frozen transformed stocks from A and B grew well on the same plates
 * C did not grow, nor did wt MF
 * yeast (or other contaminants) on two plates. This is likely from the ice bucket water.

Remaining speculations

 * wrong DNA sequence
 * too low a cell density
 * hold on ice for 10-30 minutes prior to electroporation
 * recovery wrong
 * recovery at low temperature (perhaps in the cuvette)
 * osmolarity of the recovery medium is wrong
 * Test higher osmolarity of electroporation buffer than wash medium
 * some used 0.5 M sorbitol and 0.5 M manitol, 10% glycerol
 * some buffers had Mg++, but we can't use it due to transposomes

DNA sequence analysis

 * PCR from A/B strains
 * PCR from transposome mix
 * PCR from transformed E. coli

Good controls

 * E. coli transformation
 * growth of MF tetR strain
 * measurement of CFU in transformed input


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