Richard P. Grant
see_sig at cmtech.co.delete.uk
Wed Jun 3 06:20:45 EST 1998
In the first instance, simply Taq, yes.
I've used the same idea for Pfu too. Taq I've tended to do 25 cycles, Pfu
30 cycles. Taq was a home-made buffer, Pfu (cloned) was the supplied, but
I've always titrated Mg2+ concentration.
It's a bit worrying that you're getting errors using a proof-reading
enzyme. I would suggest trying different enzymes.
Silly question - how do you know you have the mistakes? Are you comparing
with a published sequence?
In article <6l39hr$8g3 at sun0.urz.uni-heidelberg.de>,
un691cs at genius.embnet.dkfz-heidelberg.de (Clemens Suter-Crazzolara, PhD)
: I used 50 uM as well, AND a proofreading enzyme (ULTMA), so that can't be
: the problem... Did you use simply Taq ? Could you tell some more about
your conditions ?
: How many cycles for instance ?
: thanks clemens
Richard P. Grant MA DPhil | rgrant at cmtech.co.uk
Senior R&D Scientist | work: http://www.cmtech.co.uk/
Cambridge Molecular | home: http://www.avnet.co.uk/adastra
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