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
