Endy:Standard transformation positive control

From OpenWetWare
Jump to navigationJump to search

We are using pUC18 at a concentration of 1ng/uL (in sterile dionized water from the Bldg68 media room) and adding 1uL as a positive control. If you are interested in getting some of the DNA, ask Jason. A few of us will try storing our efficiency results here to collect some data on how we're doing, and identify any new appraoches that improve efficiency.

Procedure for measuring effiency

  1. Plate on LB (non-selective) to count the total number of survivors of the transformation procedure.
  2. Plate on LB+amp (selective) to count the number of successful transformants.
  • The dilution to plate will depend on the number of cells in your competent cell sample. If you know the number of cells in your competent cell sample you could also report the fraction of survivors.

Jason Results

I made a batch of TOP10 Chemically competent cells 8/1/07. Transformation with 1µl standard pUC18 yielded 9.4x10^3 transformants/50µl aliquot. -Felix