DNA ANALYSIS OF SPERM

Terry Hoy WHGTH at cardiff.ac.uk
Tue Aug 8 07:13:05 EST 1995


> We are interested in staining fish sperm with PI to be used as an
> additional ploidy marker in analyzing fish testis homogenates for DNA
> content.  Can anyone help with a successful method to preserve long-
> term (freeze) aliquots of sperm to be prepared and run along with
> other samples at a later date?  Up to this point, all methods have
> resulted in loss of cells following freezing in liquid nitrogen. 
> Also, any other hints for performing flow analysis of sperm would be
> greatly appreciated.  Thanks in advance!           Sue
> 
> Susan Dawes
> DynCorp/TAI
> USEPA Cincinnati
> DAWES.SUSAN at EPAMAIL.EPA.GOV
> voice (513)569-7721
> 

I used the Vindelov method for many years to prepare trout RBC nuclei 
for use as standards. Cytometry 3 323-327.
On one occassion a blood sample was contaminated with sperm 
following  unexpected activity from the trout! After processing,  
the preparation conveniently contained two standards, diploid 
rbc nuclei and haploid sperm. This was stable for many months and may 
prove a useful alternative method for storing fish sperm nuclei 
(alone!) for PI staining.

Terry Hoy.

Dr. Terry Hoy
Principal Research Officer
Department of Haematology                   Telephone 44 1222 743458
University of Wales College of Medicine           FAX 44 1222 744655
Heath Park                                  E-mail hoy at cardiff.ac.uk
CARDIFF CF4 4XN           http://www.cf.ac.uk/uwcm/hg/hoy/index.html
U.K.



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