User:Tk/Notebook/MF-xfm/2008/04/10

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


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Transformations

 * Made Electroporation buffer with 12% glycerol for easier freezing and more stable frozen cells
 * Transformed some fresh cells
 * 45 ml cultures, some very early (1) and some a little late (4)
 * wash 2x with 45 ml new EPB
 * shake out all liquid from 50 ml tubes
 * vortex, leaving about 250-300 ul samples
 * electroporated two samples (50 ul + 1 ul transposome), one at 1.25 KV in 1 mm cuvette, another at 1.75 KV
 * also did two E. coli controls, same conditions.
 * outgrowth in 1700 ul 1161 medium or SOC
 * Plate at 1 hour and 1.75 hours (100 ul for E. coli, 300 ul for MF)
 * at 1 hour, the growth in #2 was clearly highest, yellow culture medium. #3,4 were also somewhat grown, #1 was not


 * Realized that I had been using 1 mm cuvettes for earlier transformations, which may have been the cause of cell death and sparking.
 * No sparking this time on any samples.


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