PCR: magnesium vs annealing temperature

Ned Mantei mantei at neuro.biol.ethz.ch
Mon Jan 20 06:19:23 EST 1997


In article <853756783.8104.0 at molgennn.demon.co.uk>,
barbj at molgennn.demon.co.uk wrote:

> This is very clever but obviously you have missed the point that a lot of 
> people spend time trying many different magnesium concentrations which 
> appears to be a waste of time.

My feeling is just the opposite: We choose one temperature that seems
sensible and then try 3 or 4 different magnesium concentrations (typically
1, 1.5, 2, and 3 mM). Usually one or more of them works. This saves doing
several separate runs at different temperatures to establish the
conditions (we don't have a PCR machine that can do more than one
temperature at once).

-- 
Ned Mantei
Dept. of Neurobiology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland
mantei at neuro.biol.ethz.ch   Fax: +41-1-633-1046



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