Help with Cellular Fractionation - (Mar/13/2012 )

I'm planning to do a nuclear-cytosolic fractionation on lymphoblasts. I'd like to use hydrodynamic shearing to lyse my cells and was wondering what size needle is appropriate to break open my cells but leave the nuclei intact. I'm thinking 20-22 gauge.

Thanks for your help.

last time I used 25 gauge with a Tween 20 buffer and it didn't work. So I switched to Digitonin buffer. You can find my protocol in my earlier posts if you search my name.