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[Yeast] spores isolation

Jean-Nicolas Jasmin via yeast%40net.bio.net (by jnjasmin from gmail.com)
Fri May 22 09:59:58 EST 2009


Hi all,

I would like to isolate spores of S. cerevisiae from a mixed culture
containing spores and vegetative (haploid and some diploid) cells. I obtain
that culture by digestion with zymolyase (which, by the way, is rather
inefficient at killing vegetative cells of my strain). I am currently using
mineral oil and water: spores segregate in the oil and cells segregate in
the water. But it is almost impossible to get the spores out of the oil.
Even centrifuging at 13,000 rpm for over 20 mins won't transfer them from
the oil into water. Ultimately, my goal is to transfer the spores
(with almost no vegetative cells) into liquid growth medium. Any suggestions
on how I can isolate the spores?

I have checked the archive from the yeast list and I couldn't find a
solution to that problem.

Thank you in advance,
Jean-Nicolas
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