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PCR amplification from yeast genomic DNA

Michael Moser moser at U.WASHINGTON.EDU
Mon Aug 1 12:24:40 EST 1994


Dear Buddies,
On Sun, 31 Jul 1994, Kevin Morano wrote:

> 	The key to successful PCR amplification in our lab is Mg, Mg, Mg. 
> A given set of primers and template will do absolutely nothing at say, 3 
> mM Mg and will produce beautiful bands at 8-10 mM Mg.

My question is what if any effect does all that Mg++ have on your 
polymerase of choice?  My concern is that there could be a reduction in 
fidelity leading to an increase in sequence changes in the PCR product.

Has anybody got some experience with sequencing PCR products produced at 
such high Mg concentrations?

Mike Moser
Department of Biochemistry  SJ-70
University of Washington				  moser at u.washington.edu
Seattle, WA  98195			            Make peace my beast is yeast





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