PCR: magnesium vs annealing temperature

Dr G R Taylor grt at dial.pipex.com
Sat Jan 18 07:36:45 EST 1997

I have found that simplex PCRs are almost always capable of being 
amplified by using 50mM KCl, 10mM Tris(pH 8.8 @ RT), 200uM dNTPs and 
1.5mM Mg over a temperature range(usually between 50and 60 degrees).  But 
sometimes with multiplex PCRs you get more even amplfication of the 
different targets by tweaking Mg and temperature.  This is a pain,  but 
made much easier if you have access to a temperature gradient device like 
the Robocycler.  Then you can vary both parameters in the same 

GR Taylor PhD MRCPath
mailto:gtaylor at hgmp.mrc.ac.uk
-in vitro veritas-

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