PCR amplification from yeast genomic DNA

Kevin Morano ez005528 at rocky.ucdavis.edu
Sun Jul 31 15:26:16 EST 1994


Hi Janet,

	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. If an amplification 
is important to us, we'll usually run a Mg titration (different for each 
enzyme we try) from 1-10 mM Mg in 1 mM steps. It is of course imperative 
that primer and template concentrations remain constant throughout trials 
as variability in DNA concentrations will rapidly overwhelm these small 
Mg differences. Lastly, whatever amount of template you're using, try 
more. Good luck!


--
---------------------------------------------------------------------------
Kevin A. Morano
Internet:kamorano at ucdavis.edu    
Bitnet:kamorano at ucdavis   
Section of Microbiology, University of California, Davis



More information about the Yeast mailing list