Hello to all Thanks for all your help in the past and now I am coming back for more advice from all you awesome flow-ers. Sorry if this seems a repeat to some but I searched the previous emails and didn't find too much in the way of how to handle clumpy yeast cells but did find a lot of good information. I have a group that is working with yeast and is wishing to do cell cycle analysis on it. They have the protocol in hand for SYBR green from Current Protocols in Cytometry. They have tried the protocol once and with strange results due to clumping/possible budding. The yeast cell line, W303, becomes very clumpy through it's processing and the procedure they are working with. They are using alpha factor as the means of synchrony. I know testing out various times of sonication is needed but there is a concern that the yeast might not handle too much. Has anyone had this problem and have a solution on how to handle the clumps to get good cell cycle data if sonication doesn't do the trick? Any advice given would be greatly appreciated! I am working on an LSRII with 3 lasers, no UV. Thanks for all the tremendous support and help the community supplies! Mary *************** Mary Price Senior Research Scientist Cell Analysis Core Facility Tulane Cancer Center JBJ 455 Phone (504) 988-3422 Fax (504) 988-5516 mprice1@tulane.edu
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