DNAsing TAQ polymerase

J.J. Baker mbjjb at s-crim1.dl.ac.uk
Fri Jan 13 12:20:01 EST 1995


: >We are trying to PCR some highly conserved heat shock proteins and
: >are experiencing exceptional difficulty with PCR contamination.
: >We are aware that there are trace levels of DNA in many commercial preps
: >of TAQ and thought that it may be due to this

Are you so sure it's your Taq that's contaminated? To try and reduce the
amount of DNA in my other solutions, I set up the reaction minus taq, primers
and template and UV irradiate. I generally use a UV crosslinker setup
but a transilluminator will do but it will take longer! As a rule, i put
the reaction mix, my pippetors and tips in the machine and irradiated until
I could smell ozone! This proccess will crosslink and break DNA strands
that may contaminate your reaction but does not affect your dNTP.

I hope this helps

Jon Baker

Dept Chemistry
The School of Pharmacy
29/39 Brunswick Sq29/39 Brunswick Sq
London
WC1N 1AX



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