problems with taq-polymerase

D. KIM dkim at NMSU.Edu
Wed Jun 30 11:18:33 EST 1999


My former advisor, Dr. Paul Jackson at Los Alamos National Laboratory,
also noticed this phenomenon.  Taq polymerase preps are indeed
contaminated with bacterial DNA.  He was using enzyme from Perkin-Elmer,
and I imagine this is true for all brands of enzyme.  I do not know how he
resolved the problem.  One possible solution would be to pre-digest your
reaction mix with DNAse I before adding template and primer.  Inactivate
with heat (the Taq will be OK, of course), then add primer and template
and proceed with the run.

Daniel Kim
dkim at nmsu.edu


Eduard Post <e.post at biol.rug.nl> wrote:
: Hello,

: We have great problems with Taq-polymerase from Amersham Pharmacia. We
: are producing a pcr-product in our negative control ( no DNA added). We
: think the Taq-polymerase is contaminated with DNA, maybe from the
: organism it's isolated from    (Thermus aquaticus). We are performing a
: 16s-rDNA pcr with universal-bacterial primers.

: Are there more people with the same problem? Have you found a solution?
: Are there other polymerases (from other companies perhaps) wich have the
: same problem?

: Greetings Eduard Post

: Microbial Ecology
: Rijksuniversiteit Groningen

: E.post at biol.rug.nl




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