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
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.
Nick