disappearing DNA

O'Donohue odonohue at lille.inra.fr
Fri Dec 5 05:36:19 EST 1997


Has anyone had trouble with disappearing DNA inserts? We have isolated a
gene from an archaebacterium by PCR and have tried to clone the DNA into
various vectors (pGEM-T, pZero II, pUC18). However, we don't get many
colonies after transformation and the ones we do get give strange
results: often on dot blot analysis several clones light up but when we
analyse the plasmid DNA either the insert is missing,or is smaller than
expected, or worse, not only is there no insert,but bits of the plasmid
are missing too. Can any suggest a way around this instability problem?

Thanks in advance.

Michael J. O'Donohue
odonohue at lille.inra.fr



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