How to crack yeast for PCR ??

Jeremy Marcus marcusj at helix.mgh.harvard.edu
Wed Apr 10 18:57:50 EST 1996


In article <liss.15.316BD4A9 at alf1.ngate.uni-regensburg.de>,
liss at alf1.ngate.uni-regensburg.de (Michael Liss t4310) wrote:

_snip_

< Let me come to my question:
< Is this DNA-isolation really necessary or is there an easier way to
< perform a reliable PCR. (Just like with E. coli where you just take a 
< little colony and add the whole cells to your reaction.)
< If anybody has experience with this or can contribute any other 
< suggestions on how to run the PCR etc. please let me know...

Mike,

I've known people to routinely PCR amplify from plasmids in yeast using
the same method as you describe above for coli -- just add a little of the
yeast to the reaction and the initial denaturing cycle presumably bursts
the yeast.

I tried it myself once, but got extremely variable results.  It worked the
way I expected about 25% of the time, but, obviously, that's not good
enough.  Perhaps others have some additional ideas about how to enhance
the efficiency?

Jeremy



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