PCR screening: False positives
Dr. W. Greenhale
wgreenha at hgmp.mrc.ac.uk
Tue Sep 6 11:38:57 EST 1994
Hi, a colleague of mine recently came across what I
considered to be a fairly worrying phenomenon.
We routinely PCR screen E.coli colonies in our lab
and sometimes find PCR positives which are
\expression negative (not very surprising). However,
it seemed that this was more common after
electroporation. Acting on a hunch my colleague
therefore tried PCR-ing (I take it the verb to PCR
is now official English grammar at least on the bionet)
up random areas of colony free regions on her plates
and came up with a remarkably high level of PCR
positives. Obviously the reason is that traces of the
transforming DNA were present on the plate. This can
be overcome by carefully picking the centre of colonies
(not always easy especially in a big screen). What I
was wondering is if this problem is common knowledge
and if not shouldn't people be warned. I for one had total
faith in the PCR screening technique and have almost
certainly wasted quite extensive periods expressing
negatives.
Bill Greenhalf
MRIC Biotech.
NEWI - Deeside,
Connah's Quay
Clwyd, UK
E.mail Greenhalfw at newi.ac.uk
or as above
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