DNA extraction from bone
haroldb786 at aol.com
Sat Sep 6 19:09:12 EST 1997
First try organic extraction (phenol/chloroform), if that doesn't work,
then you may have to use a silica-based extraction...... either from
companies (the names of which escapes me right now) or make your own. I
worked on silica extraction of 1000 year old whale bones with some
success.... I would search www.biotechniques.com for articles, or download
the procedure I help adapt and is now part of the Fish and Wildlife
Forensic Lab at http://ash.lab.r1.fws.gov./labweb/for-lab.htm under
protocols--- you will need the acrobat reader software-- also available
Part of the work all depends on what type and amount of bone material you
have access to........ You will need to grind the bone into a powder---
liquid nitrogen and a morter/pestle and extract 0.5 grams...
The other question is what part of the genome are you looking at??? Old
bones tend to have very degraded DNA.... So, you are only going to be able
to amplify 200-400bp... With the whale work, I designed primers for a
150bp fragment in the mitochondrial cytochrome b region. This circular
genome is relatively durable to degradation..... and there usually more of
it than nuclear DNA.
You should also contact Steve Fain at the Forensic Lab (he is the person
to taught me everything) at: steve_fain at mail.fws... explain what you are
trying to do.....
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