[Yeast] Re: Sporulation/tetrad defects

[eileen.mitchell at syngenta.com]

Hello all,

I, too, am having sporulation frustrations!  I am using an S288C
background strain and have mated some deletion haploids and selected for
diploids.  Then I have made glycerol stocks by patching isolated
colonies onto YPD.  Then I have use the same method as the Saccharomyces
Genome Deletion Project, from Linda Riles and Matt Curtiss:
http://www-sequence.stanford.edu/group/yeast_deletion_project/spo_riles.

I managed to get the method to work last year, but now even the control
diploid (with no deletions) won't even work.  I have tried leaving at v
low temperature as a colleague told me that would work (15 degrees C).
I have also tried patching on 1% potassium acetate + supplements but no
joy there yet, either.  I tend to get a number of 3-spored asci for the
control and maybe one 4-spored ascus on a good week, but no efficiencies
high enough to dissect.
Can anyone give me any advice?  I don't understand why it used to work
and has now stopped - I don't think I have inadvertently selected for
non-sporulators as I make fresh plates from freezer stocks every time.

Thanks,
Eileen

Eileen.mitchell at syngenta.com

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