Electroporation advice needed

Ian Mehr ijmehr at merle.acns.nwu.edu
Tue Apr 25 13:51:34 EST 1995


In article <3n7scq$lmu at ousrvr.oulu.fi>, jkortes at paju.oulu.fi (Jarkko Kortesmaa) says:
>
>I recently tried transforming E.coli by electroporation. Efficiency was 
>not very high, only 1,5x10^6.


Three things come to mind:  when resusspended in 10% glycerol after washing, the cells
should be at a concentration of 10^11 per ml.  Any less will result in logs lower
efficiencies.  Furthermore, the DNA added must be very clean, and free of salts.
The DNA should not be suspended in more than 0.5x TE.  Lastly, after electroporating,
the samples must be _immediately_ added to growth media.  Hesitation of only one
minute can result in a log or two lowered efficiency.

Ian J. Mehr
ijmehr at merle.acns.nwu.edu



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