Plasmid losing insert??

Dennis O'Neill doneill at WOLF.BIOENERGY.COM
Thu Apr 4 13:24:18 EST 1996

Sorry to ask what is probably an introductory microbiology question. What
would be probable causes for our pNoTA plasmid to loose its insert yet
still remain viable? Ampicillin concentration is fine and not too many
freeze-thaw cycles have been done, so what other variables could be
responsible for this? Restriction digests and ScreenTest have been done,
plasmid is viable but insert in mostly present in a small number of cells,
by PCR results. The insert size is only 2 KB and not just one insert has
been problematic??

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