lysing S. cerevisiae cells

Debbie Nathan dnathan at
Thu Jan 13 14:45:55 EST 1994

We are having trouble making good native S. cerevisiae lysates. 
Generally (in the cold room) we beat a concentrated cell slurry with an
equal volume of 0.45 uM glass beads for 5 x 30 sec using a hand vortexer
set at the highest speed or for 2 min using a VWR multi vortexer.  While
we don't have much of a problem with protein degradation using this
method, our yields of protein are quite low.  When we have tried
increasing the time we beat the cells, we get degradation.  Does anyone
have a breakage method they really like for native lysates? Thanks.
Debbie Nathan

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