DNA from old bones

Peter J. Snow snow-peter at yale.edu
Fri Jun 25 13:22:41 EST 1993


In article <9306251536.AA14638 at pooh.UCSD.EDU> andy at POOH.UCSD.EDU (Andy Dizon) writes:
>
>We have a need to amplify DNA from museum skull specimens.  When there is 
>skin tissue stuck on the old bones, we expect we won't have any trouble.
>However, what about cleaned bones?  We'd appreciate any hints, references,
>warnings (old curses, etc.) regarding extraction methods.
>
>Andrew
>
>Southwest Fisheries Center
>La Jolla CA 92038
>FAX 619 546-7003



Andrew,
	About three years ago, I did some work with Bison bones and teeth
that were about 130 years old. I broke up the bones and teeth into small
fragments with a hammer, and then soaked them in homogenization buffer for
a few days on a shaker/rotating platform. I then just followed a normal
DNA extraction protocol. I also tried just splitting open the limb bones
and taking scrapings from where the marrow used to be. All methods yielded
large amounts of partially degraded DNA. However the greatest amounts were
recovered from teeth that were still in their sockets. Work on other bones
(e.g. goat limb bones found out in the desert) also led to good amounts of
DNA. PCR of mitochondrial (I never tried nuclear) DNA worked well most of
the time with this material. If you can get well preserved teeth though,
use those.  

         One thing to watch for is when you run your extracted DNA out
on an agarose gel. There are often some altered sugars that will fluoresce
under UV and give a false impression as to how much DNA is there. It is a
good idea to take a photo of your gel both before and after you soak it in
EtBr.

	 As regards curses, well my thesis (on a different topic) has not
really gone well since....and as for that blood that appears on my
walls every night at midnight.....

Peter Snow, 
Biology Dept.,
Yale University.

e-mail: peter at doliolum.biology.yale.edu



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