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SDS Precipitation with 1% Lysis Buffer after 10' on ice - (Apr/30/2007 )

I have seen one post entry about this topic but no reply. Hoping for better odds this time around.

Every time I incubate my cell pellet with 200 ul of 1% SDS lysis buffer (Upstate), the SDS precipitates out. Before I sonicate I always have to warm up the pellet with my hands to get everything back in suspension.

I was wondering:
1. Should I even worry about the SDS precipitating out b/c the sonication is likely to warm up the sample anyways?

2. Or if it is a problem I should be concerned about - is there a different way I should be incubating (like in on ice water)?

Thanks in advace for the response (hopefully).


-lauren Czaplicki-

Hi Lauren,

the best thing to do is to perform an experiment on it.

When I did ChIP I did warm up the suspension to get the SDS back into solution, I have not tried just going ahead with the sonication. Having said this, I sonicate in an ice bath so I can't really see if the SDS precipitates again once in the ice bath, it's probably a likely thing to happen though.