Help with Genomic PCR

Miguel Cabrita mcabrita at gpu4.srv.ualberta.ca
Wed Aug 21 20:19:54 EST 1996


Hi, I am interested in hearing from people who have successfully used PCR
to amplify large regions of DNA.  I have a gene subcloned into
pBluescript.  The insert is in the region of 12kb.  I have a variety of
primers based on the cDNA sequence which I would like to use to do a quick
analysis of the gene structure but this may involve PCRing several kbs at
a time which Taq is unhappy to do.  I know there are various kits and
variations of enzymes on the market from various companies.  Can anyone
suggest what to use and what not to use?  Thanks
Imogen Coe, Ph.D.
Dept. Biochemistry
University of Alberta
Edmonton, AB
Canada
icoe at gpu.srv.ualberta.ca



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