Yeast Colony PCR

Simon Twigger simont at post.its.mcw.edu
Thu Jan 25 12:03:27 EST 1996


I am using the Clontech 2 hybrid system and its come time to check my 
postitive colonies. The manual suggests doing a quick/crude plasmid prep 
for each clone, then PCRing off that and then restricting the PCR product 
using AluI and/or HaeIII to try and identify similar clones - thereby 
cutting down on the amount of sequencing to be done.

I was wondering if there is a protocol for Yeast Colony PCR, like the one 
we regularly use with bacteria, so that I can PCR my product directly 
from a colony without having to go through the plasmid step? I have heard 
that the problem tends to be getting the yeast open and also the genomic 
DNA contamination. 

Has anyone tried this/published anything on this anywhere?

Thanks for any help anyone can give me

Simon Twigger,
Medical College of Wisconsin, Milwaukee.




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