glass beads for yeast lysis

[Tomas Drgon]

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)