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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