genomic DNA PCR

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Wed Oct 4 10:06:58 EST 2000


In article <8rf9jl$n0h$1 at hpngsv01.mgh.harvard.edu>, Robert Mihalek
<rmihalek at partners.org> writes
>Can someone forward me a general protocol for doing PCR using genomic
>DNA as the template?  I seem to recall needing about 100 ng of DNA, but
>I forget stuff like  how much Taq to use, concentration of dNTP's, and
>concentration of primers.

Buffer
1.25u Taq per 50ul PCR (add last)
MgCl2 1.5mM
200uM dNTPs
0.5uM each primer
30ng human xsomal (10,000copies)
water to 50ul

Duncan 
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNamp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk






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