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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MF-xfm
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