Isolation of nulcei

[Kristina Warton]

Hello,

I'm looking for a protocol for isolating nuclei from cultured human 
and rodent cells. I'm after a protocol which does not use detergents, 
so that the membranes of the nuclear envelope remain intact. The 
protocols I have been using either leave most of the cells 
undisrupted, or cause the nuclei to clump. I'm worried that spinning 
clumps of nuclei through sucrose to purify them will not do anything 
towards removing the cell debris around individual nulcei.

Any ideas ?