glass beads for yeast lysis

BRAINSL at aol.com BRAINSL at aol.com
Mon Sep 25 07:56:34 EST 1995


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In a message dated 95-09-20 11:28:41 EDT, BIOSCI-REQUEST at net.bio.net writes:

>Subj:	RE: glass beads for yeast lysis
>Date:	95-09-20 11:28:41 EDT
>From:	tomasd at bdg10.niddk.nih.gov (Tomas Drgon)
>Sender:	postman at alw.nih.gov (AMDS Postmaster)
>Resent-from:	BIOSCI-REQUEST at net.bio.net
>To:	yeast at net.bio.net
>
>In article <bicfo.14.305E5AC7 at flinders.edu.au>, bicfo at flinders.edu.au wrote:
>
>> hallo all!
>> Can someone give me the good oil on using glass beads to lyse yeast? I'm
>using 
>> 1ml of lysis buffer (50mM HEPES, pH 7.4, 150mM NaCl, 5mM EGTA, 5mM EDTA,
2%
>
>> Triton x-100 and inhibitors) to ~ 6 x 10^8 cells and 1.5g 400 -600u 
>> acid-washed beads. I'm vortexing them to death - and yet I'm getting zilch
>in 
>> the way of lysis! What am I failing to do?
>> Replies gratefully received!
>> Cheers, Colleen
>
>Hi. 
>I use simply 0.05 M Tris/HCl pH 7.5. No detergent, no salt, nothing. I use
>about 0.5 g of wet cells in 2 ml breakage buffer. Add beads so that there
>is still about 4 mm of free liquid above the beads, and vortex 30''/30''
>on ice (10x). At that point they are totally dead, you can barely find a
>living cell in micro.
>
>Tomas
>
>-- 
>Vidno jak psoj kulky.
>(Old Slovak proverb)
>
>
>
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>Date: Wed, 20 Sep 1995 14:06:51 GMT
>From: tomasd at bdg10.niddk.nih.gov (Tomas Drgon)
>Subject: RE: glass beads for yeast lysis
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