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

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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.