Synchronization

Robert J.D. Reid reid at hendrix.jci.tju.edu
Tue Apr 28 10:38:21 EST 1998


Dear Yeast group,

  I am having trouble with flow cytometry after alpha factor 
synchronization.  A significant number of the G1 cells after alpha arrest 
have gone through cytokinesis but remain stuck to their previous bud.  I 
sonicated these before flow cytometry, but this did not disrupt the 
pairs.  My flow cytometry profile shows two sets of G1 and G2/M peaks 
through the first and second cell cycles.  I can gate out the second set 
of peaks, but it is a significant fraction of the population and it 
varies somewhat by sample.  Any ideas about breaking up these pairs?  I 
am going to try a light zymolyase treatment and perhaps more sonication, 
but if anyone has any ideas or more specific protocols I would appreciate 
the advice.

Thanks,

Bob.


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