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