RNA extraction from formaldehide fixed cells - (Sep/25/2006 )

hi everybody,

I want to fix cells with formaldehide, permeabilize them, incubate them with an antibody and sort them, and then extract RNA from sorted cells. is it possible? any protocol? should I reverse crosslink or something?

thanks a lot

if i got your exp, you wxant to do kind of cell sorting by antibody mediated way, and then extract RNA?
But i think the step is not that good because of the alteration of global RNA induced by formaldehyde, and by the possibility of RNAses to access RNA.

What i would suggest is to use RNAlater during your selecting period (if feasible) and then extract RNA in liquid way (not by columns i mean, so use trireagent or trizol).
How is done the reverse cross linking?
I think you need to reverse the cross link for having a good RNA analysis possibility, as even in denaturing buffers, the RNA would not be "un cross linked"

hi fred, thanks for your answer. yes, that's want i want to do. about the reversion of crosslinking, i thought that maybe it is possible to apply the same protocol (or modified in some way) as for DNA in ChIP assays, but i don't know if that can be done with RNA.

thanks again