Yeast PCR Advice

killer yeast vvsvetlov at utmem1.utmem.edu
Wed Nov 15 01:32:52 EST 1995


In article <1995Nov2.091333.1 at mcvax2.d48.lilly.com>,
books at mcvax2.d48.lilly.com wrote:

> We are using a yeast two-hybrid system and would like to PCR the
positive yeast
> clones. I am told the yeast cell wall is diffcult to break open. Is there a
> yeast PCR prcedure or does anyone have advise for doing PCR with yeast? Thanks
> DPSMITH internet:smith_dennis_p at lilly.com
Filamentous fungi cell wall is hard to break (Ha!). Yeast is a dolly. What
we do for PCR in yeast is to use rapid plasmid DNA release protocol
(available in CPMB and white book - let me know if you can't find it) with
glass beads. Entire procedure takes about 30-45 min and lives you with
PCR-ready DNA. 
Have fun, 
Vladmeister

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