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RNA extraction

Bernard Murray bpmurray*STUFFER* at socrates.ucsf.edu
Tue May 12 18:17:40 EST 1998


In article <35579EBF.7F0F at ucla.edu>, Curt Eckhert <ceckhert at ucla.edu> wrote:

> We want to extract total RNA from yeast using Quigen RNeasy Midi kits,
> but we don't have a bead mill or zymolase.  However, we do have a
> sonicator.  Can I use the sonicator for the intial mechanical
> disruption?  Has anybody done anything like this before?  Does anybody
> have any other methods of mechanical disruption?

As someone very new to yeast handling this is a question very dear
to my heart.  I don't think you need a bead mill unless you are trying
for heroic quantities of RNA or need to process lots of samples.
I've had good success so far using silica/zirconia beads and just
vortexing them.  Being somewhat paranoid about RNases I vortex in
a home made version of Trizol and then finish the extraction with
hot acid phenol.  Using this method I generally get plenty of RNA
from 20 ml of mid/late exponential culture.

If there are any other effective methods of RNA extraction from
yeast I would also be glad to hear of them.
   I hope that this helps,
      Bernard

-- 
Bernard Murray, PhD
Dept. Cell. Mol Pharmacol., UCSF, San Francisco, USA






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