>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)
In the Methods in Enzymology Yeast book (Guthrie & Fink, eds) there is an
article by (I believe) Beth Rockmill and Shirleen Roeder. In it they
describe an ether vapor method that selectively kills vegetative cells. I
have used it and it works.
Michael Lichten
lichten at helix.nih.gov
