Subcloning dirty PCR products

Bernard Heymann bheymann at bragg.bio.purdue.edu
Sat Sep 3 12:14:31 EST 1994


In article <34a0ilINN1k8f at sat.ipp-garching.mpg.de>,
krasel at alf.biochem.mpg.de (Cornelius Krasel) wrote:

> The hypothesis offered by the authors is that the Taq polymerase sticks
> strongly to the DNA. Since agarose electrophoresis is not denaturing for
> proteins you would expect the polymerase to co-migrate with the DNA.
> Even denaturing of proteins with phenol seems to leave Taq polymerase
> on the DNA.

I routinely purify my PCR fragments with diatomous earth in 4M GuSCN before
restriction with reasonable success. I always assumed that either Taq pol
would be denatured or at least removed from the fragment. Is there anybody
out there that have some information about this?

-- 
Bernard Heymann
bheymann at bragg.bio.purdue.edu



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