zygote germination

Dave Mitchell MITCHELD at VAX.CS.HSCSYR.EDU
Thu Jun 29 08:30:02 EST 1995


Robert C. Hodson writes:

 There are times in classical genetic analysis when it would be helpful to be 
 able to dissociate zygote clumps before spreading on germination 
 plates.  Treatment with proteinase K does not work.   Does anyone have an 
 idea or success in this regard? 

Maybe I'm missing something about the way you want to germinate your zygotes, 
but the simple answer is, don't let them clump to begin with!  We spread 
mating mixtures at low density, 1-3 hr after mixing gametes, onto complete 
(minimal) medium.  After 24 hr, plates are wrapped and left dark until 
germination time.  If the mating mixture is streaked along one edge of the 
plate, or in a square, then tetrads can be dissected directly onto the unused 
portions of the plate.  If the mixture is evenly spread on the plate at an 
appropriate density, then germination of a chloroformed plate results in 
a plate of several hundred individual zygote colonies.

David R. Mitchell
Dept. Anatomy and Cell Biol.
SUNY Health Science Center
Syracuse, NY
MITCHELD at VAX.CS.HSCSYR.EDU




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