S. epi electroporation

Kevin Nawotka nawotka at earthlink.net
Wed Dec 3 18:27:49 EST 1997


   I have been able to sucessfully electroporate S. aureus with high
efficency (almost 10*6 /microgram with pLI50). I have made S.epidermidis
electrocompetent by 2 different methods  but can't get any transformants.
The plasmid was passaged through RN4220 in all cases.

   One method uses 0.5 M sucrose as with S.aureus, and electroporation
conditions of 2.0 KV, 25 microfarads and 100 ohms.

   The other uses 10% glycerol but at room temp and the same
electroporation conditions.

Does someone have a sure fire method  to electroporate S. epi out there?

Also, is there a difference in the tolerance of various species of Staph to
chloramphenicol? I used 15 micrograms/ml to select for transformants with
aureus. I am trying another electopration with 5 micrograms/ml as a
selection to see if it makes a difference.

TIA

Kevin Nawotka



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