Yeast PCR Advice

zhuang zuo zuoz at phibred.com
Wed Nov 29 18:19:23 EST 1995


Hamid_Khoja at cc.chiron.com wrote:

>In article <vvsvetlov-1511951432520001 at tcooper2.utmem.edu> vvsvetlov at utmem1.utmem.edu (killer yeast) writes:
>>From: vvsvetlov at utmem1.utmem.edu (killer yeast)
>>Subject: Re: Yeast PCR Advice
>>Date: Wed, 15 Nov 1995 14:32:52 +0800

>>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

>>-- 

>Look in the November isuue if Biotechniques.  There is a simple method that 
>does not involve plasmid isolation prior to PCR.


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I've PCR'ed genomic targets from S. pombe and S. cerevisiae by the
simple expedient of picking a small amount of cells from a colony and
twirling it in the PCR reaction mix.  I can't imagine it wouldn't work
from plasmids in yeast as well.

John 
mcelverja at phibred.com




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