Protocol for PCR straight from yeast colony?

Steve Oysdotun koyedotu at gpu.srv.ualberta.ca
Sat Apr 8 13:21:10 EST 1995


Hi! I have done PCR with whole yeast cells before. It is basically the
same way as you do with DNA template. Only thing you should keep in mind
is that you should try to use specific primers and try PCR at high Tm
(which means your primers will be better over 20-mer). other than that you
may want to wash the yeast cells before PCR reaction. In addition, you
should include enough yeast cells (transformants) otherwise, you won't get
enough DNA fragment to be visualized on gel with EtBr staining. Besides,
bring negative controls which may apply. Good Luck!


> I need to verify if the plasmids that I transformed into my S. cerevisae
> are correct.  I know that it is possible to pick a colony of yeast, lyse
> it, and PCR amplify a fragment of DNA using the DNA retrieved from the
> yeast colony as the template.  However, I do not have a protocol for this
> procedure.  If anyone knows a protocol or where to find it please let me
> know. 
 John Stebbins



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