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Freezing Lysates before Sonication - (Jan/23/2007 )

Has anyone ever stopped the ChIP protocol after resuspending their cells in lysis buffer by freezing the lysates and sonicating at a later date? If so, did it decrease sonication efficiency or even ruin the whole ChIP?

Thanks in advance.

-lauren Czaplicki-

QUOTE (lauren Czaplicki @ Jan 23 2007, 09:47 AM)
Has anyone ever stopped the ChIP protocol after resuspending their cells in lysis buffer by freezing the lysates and sonicating at a later date? If so, did it decrease sonication efficiency or even ruin the whole ChIP?

Thanks in advance.


I always freeze the pellet of the sonicated lysate but I don't think it's to much of a problem if you froze the resuspended cells (as long as you resuspended in buffer containing protease inhibitors). The resuspension should be enough to lyse the cells and allow the inhibitors to block proteolysis. I really don't think it would have an effect on sonication efficiency.

Joel

-KPDE-