PCR mistakes

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?

R

In article <6l39hr$8g3 at sun0.urz.uni-heidelberg.de>,
un691cs at genius.embnet.dkfz-heidelberg.de (Clemens Suter-Crazzolara, PhD)
wrote:

: 
: 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
      -- Standard corporate disclaimers apply --



More information about the Methods mailing list