extractin mtDNA from fish scales

Michael Gatt mgatt at uoguelph.ca
Mon Jan 15 15:35:11 EST 1996

I am trying to extract mtDNA from walleye scales in sufficient quantity 
and quality which can be used in a Polymerase Chain Reaction.  So far I 
have used a standard DNA extraction protocol that involves using 6-8 scales 
incubated over night at 60 C in Proteinase K and buffer 10mM Tris ph 8.3, 
50mM KCl, 0.5% Tween 20.  The samples were then heated at 95C to 
inactivate the remaining proteinase K.  After that I PCRed as I would 
with other mtDNA samples extracted from tissue.  My control sample worked 
but the DNA from the scales didn't.  I am not sure whether I need to try 
phenol-chloroforming (it has been recommended that I don't do this) or 
whether this would reduce the small concentration of DNA already 
present.  Any information on other ways that I may get this to work would 
be appreciated.  Or maybe one will have recommendations on ways to 
inprove the present technique that I used.  

Mike Gatt
Department of Zoology
University of Guelph
Guelph, Canada

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