Yeast PCR Advice
Hamid_Khoja at cc.chiron.com
Hamid_Khoja at cc.chiron.com
Wed Nov 15 13:21:34 EST 1995
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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