How can I disrupt ascospores of yeast?

Colin Rasmussen colin at pombe.usask.ca
Mon Feb 7 10:22:16 EST 2000



"Tim [iso-8859-1] Söllick" wrote:

> I am looking for a protocol to handle spores after I have mated two
> strains of yeast and induced sporulation.
> How can I get rid of remaining zygotes? How can I disrupt the spores to
> grow them on selective medium?

Standard procedure is to take some of the mating mix and incubate with
glucuronidase overnight in water (filter sterilize before adding cells).  This
digests the cell walls of vegetative cells but leaves the spores intact.  Once
you done that, collect the spores by centrifugation, wash a couple of times
with sterile water.  Then plate a series of diluted samples to determine viable
spore concentration.  As far as disrupting, just grow them on rich medium
first, then replica plate on whatever selective media you wish to test.

Colin



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