User:Tk/Notebook/MF-xfm/2008/04/16

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Transformants

  • One new transformant on the no-DNA no-BSA plate
  • The BSA and DNA transformant from Friday grew cultures in tet medium
  • No new transformants from Monday (none expected)
  • E. coli transformation showed 62 colonies, down from previously
    • probably because I electroporated at 1.5 KV rather than 1.75 KV

New tranformation tests

  • transform testing 4x and 40x concentrations with EPB (no glycerol)
  • three washes at 45 ml each
  • resuspend 50 ul of the 40x concentration in 450 ul for the 4x concentration
  • aliquot 3x 50 ul of low and high concentrations
  • Add 0, 10 ul, 30 ul of 1 M sorbitol as an adjustment to osmolarity
  • Electroporate at 1.5 KV
  • outgrowth for 1 hour at 30C
  • plate 300 ul
  • E. coli transformants done at 1.5 KV and 1.75 KV