PCR on cells to identify transfectants?

Markus Winter beyonder at blueyonder.co.uk
Thu Aug 3 15:45:45 EST 2006


The idea is to save on expensive reagents.

The selected clones have an intact Kan/Neo cassette (otherwise they
would not survive).

Given that the construct could have opened anywhere else in the plasmid
for integration there is a 20% chance that the incorporated construct
has an intact expression cassette with my protein (probably much less
if the protein proves to be toxic).

So I want to first check by PCR if the expression cassette is intact in
the genome.

Given what can be done in forensics nowadays I thought there must be a
quick and dirty way ...

Markus



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