Help with Genomic PCR

Taek H. You tyou at postbox.acs.ohio-state.edu
Sat Aug 24 01:09:11 EST 1996


In article <4vgcjq$gcq at pulp.ucs.ualberta.ca> mcabrita at gpu4.srv.ualberta.ca (Miguel Cabrita) writes:

>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

Try the paper by Barns in PNAS (1994, Feb. or Apr.).
It will explain what's going on long PCR, and you will get some idea what you 
should do and what you should choose.
Sorry that I don't have the ref. on hand with me right now.



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