nuclei isolation

From: Neil Young (n.young@aims.gov.au)
Date: Sun Oct 05 1997 - 19:37:44 EST


I was wondering if anyone had some advice on the best protocol to use for
nuclei isolation from oocytes.  I am using prawns with eggs of 200-300
microns and nuclei of approx 100 microns.
Also if there is any literature which they think is pertinent could they
passs it on.  Most of the literature I have found concerns nuclei isolation
from either cell cultures or tissues.  
The eggs I am using were stored at -80 degrees after snap freezing with
liquid nitrogen.
I don't think simply using detergents will break the egg membrane to
release the nuclei.  At the moment I am using  a glass homogeniser (0.15 mm
clearance) to disrupt the eggs prior to attempts to isolate the nuclei from
the egg shells and other debris.
Any help would be greatly appreciated



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