I am working with the plasmid pBT20, which is amp and gent resistant. I was recently told that pBT20 should only be inoculated onto LB Amp 100 agar, and never cultivated on LB Gent or any other Gent media for overnight cultures. Why can pBT20 only be grown on amp100 and not gent?
Michael J. Benedik
I believe that the gent marker is on the transposon of pBT20. So if you select for Gent resistance you might just select strains that carry a transposition event and not the actual plasmid itself. So it is safer to select for amp and be sure you are actually maintaining the plasmid.
Having said that, the addition of gent is not likely to be deleterious and probably would work fine, but amp is a better selection marker for the plasmid.
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