I am trying to isolate some random spores from a poor sporulating diploid.
The problem that I am trying to avoid is the propagation of the non-sporu-
lated diploids. So far in the sporulation mixture I have two, three, and
four spored asci. I would like to focus on these and eliminate any
non-sporulated diploids. I have used a method described in Methods in
Enzymology which calls for the use of 2% potassium acetate rather than
1%. This is supposed to kill of any vegetatively growing cells, but
when I observe the sporulation mixture I see normal looking non-sporu-
lated cells (in rather high proportion). Are these cells viable after
normal sporulation or 2% potassium acetate growth? Does anyone have
a method for ridding the population of non-sporulated cells?
Thanks in advance,
-Al (aplummer at sprint.uccs.edu)
