Removal of Non-sporulated Diploids

AL R. PLUMMER aplummer at sprint.uccs.edu
Mon Jan 2 14:28:50 EST 1995


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)



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