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Axolotl spermatophores

MCLEANM at VAX.CS.HSCSYR.EDU MCLEANM at VAX.CS.HSCSYR.EDU
Mon Feb 17 09:03:27 EST 1997


I am trying to design an axolotl transgenic by using a restriction enzyme
mediated insertion of DNA into sperm nuclei.  Now the protocol that is 
used in Xenopus requires sacrificing the frog and though periodically we do
sacrifice 1 or 2 for tissue, I may need more sperm than what that would give.

So, being the males lay the nice spermatophore cones I want to try to isolate
the nuclei from there.  The only problem I am having is getting rid of the 
extra gel coat of the cones from the sperm itself.  After spinning the tips of
the cones down and several washes etc I can't get rid of the gel.  I have not
yet tried a 2.5% cysteine solution that is used to chemically dejelly Xenopus
eggs but will be trying that soon.

I was wondering if you knew of any method to separate the sperm from the jelly
cones they are laid in?  I need to remove this before I can demembranate the
sperm to isolate the nuclei.

Thank you for your time and suggestions.

Sincerely,
Matt McLean

MCLEANM at VAX.CS.HSCSYR.EDU




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