Spore germination on YEPGal

Daniel Zabetakis dan at cubmol.bio.columbia.edu
Fri Mar 4 12:07:52 EST 1994


In article <CLzp15.8n9 at discus.technion.ac.il>,
Andrey Tsouladze <tsoul at techunix.technion.ac.il> wrote:
>
>Hi everybody,
>
>Recently, I was making a tetrad analysis in which some haploid progeny
>was to be kept alive only due to the presence of a gene under the GAL
>promoter control. Therefore, I put dissected tetrads on YEPGal plates. To
>my surprise, no single spore appeared to grow after four days. Does
>anybody have an idea why? Is it possible that glucose substitution with
>galactose prevents germination? What can I do to avoid the problem?
>

   It sounds to me that you are actually killing the spores during
dissection. This can be for a number of causes. It's hard to imagine
a diploid genotype that would give you no spores.
  People in my lab have done germination on galactose. You might expect
an increase in inviability (we have found that our strains give a lot
of gal- mutants), but you shouldn't get zero.
   Do a test dissection on an isogenic strain that doesn't depend on your
plasmid.

DanZ

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