nuclei isolation
Neil Young (n.young@aims.gov.au)
Mon, 06 Oct 1997 10:37:44 +1000
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