PCR from yeast genomic DNA

Richard Stevens rstevens at U.WASHINGTON.EDU
Mon Jan 8 14:14:09 EST 1996


Hey Jon,
     If you have access to live Saccharomyces cultures or colonies, you 
may be able to PCR amplify directly from the colony.  People in my lab 
have no trouble just adding a yeast colony to the PCR reaction and 
getting a product.  It is important to add mineral oil to the reaction 
even if you have one of those new-fangled thermocyclers because the oil 
seems to remove some inhibitory agent from the PCR reaction.  It is also 
important to use a new toothpick or flame sterilized loop to add the 
yeast colony, or you may get contaminants.

Rick Stevens
Department of Biochemistry
Box 357350
University of Washington
Seattle, WA 98195


On 8 Jan 1996, Jon Stewart wrote:

> I need to PCR amplify two Saccharomyces genes of approx. 1000 bp, 
> neither of which contains an intron according to the sequence database.  
> Since I have never attempted PCR from yeast genomic DNA, I have some 
> questions.  Is it possible to use the S. cerevisiae genomic DNA (> 100 
> kb) sold by Promega ($95 / 100 ug) for this purpose?  My other simple 
> choice is the Clontech Quick cDNA ($310 / 20 reactions).  Obviously, if 
> the Promega genomic DNA will work, I can find other uses for the $200 
> price difference :-).  In addition, any hints or special tricks for 
> running these PCR reactions would be greatly appreciated.
> 
> Jon Stewart
> Department of Chemistry
> University of Florida
> 
> 
> 
> 
> 



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