re yeast colony PCR

Robert Preston rapr at MED.PITT.EDU
Tue Jul 5 21:03:16 EST 1994




> Has anyone done genomic PCR with isolated yeast colonies? If so, Could you
> give me info. on the protocol. I tried to do it with colonies of Pichia
> pastoris. I followed the same procedure as with bacterial colony PCR, i.e.
> boiling cells in PCR buffer + dNTPs+ primers at 100C for10 min, quick spinning
> followed by addition of taq polymerase. This procedure did not work for even
> my positive control. Thanks a lot for your response.
> 
I've heard that that sort of thing works only if you use an oil overlay
rather than an oil-less machine, (with yeasts, I mean); SO, are you using
oil or not?? please post answer on the bboard.

Rob
rapr at net.bio.net




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