RE: Extracting DNA from bone

From:"Su, Phy-Huynh" <psu@shctampa.usf.edu>

We mince fresh bone or cartilage tissues in PBS finely first, with razor
blades held by a hemostat, or by #20 scalpel.  Remove the PBS as much as
possible, pool the minced tissues in a 50ml POLYPROPYLENE tube (weigh them
if you need to at this point), and freeze the tissues in tube in liquid
nitrogen (polysterene tube will break). 
Next, we pool  a little bit of tissues (sometimes it's very hard to scrap
tissues out from the tubes, so we smash and break the bottom of the tube
with a hammer) into a chilled, metal puvelrizer (?) in a dry ice box.  Then
using a hammer, we smash the tissues manually (requires a lot of muscle) to
powder, and remove the powder with a chilled spatula into chilled tubes.  At
this point, you can keep the tissues frozen (-80 degree C) if running late.

Another way (better, but need instrument):  using a magnetic pulverizer in a
liquid nitrogen bath.  From the wet, weighed tissues, we can put tissues in
pulverizing tubes, immerse into liquid nitrogen, and pulverize.  This is
better because the powder is much finer, requires less strength, and faster.
But you need the instrument.
Finally, we use a tissue homogenizer to homogenize tissues in extracting
buffer, either for DNA, RNA, or protein.
If you need details, tricks here and there, give me a call.  I'll be happy
to help.
Su


> -----Original Message-----
> From:	cmcbride@tulane.edu [SMTP:cmcbride@tulane.edu]
> Sent:	Tuesday, February 06, 2001 10:13 AM
> To:	histonet
> Subject:	Extracting DNA from bone
> 
> Hi,
> Does anyone know how to grind bone into a fine enough powder to extract
> DNA 
> from it?
> Thanks,
> Christine
> 
> Christine McBride, M.S.
> Tulane University Medical Center
> Center for Gene Therapy
> SL-99
> 1430 Tulane Ave.
> New Orleans, LA 70112
> cmcbride@tulane.edu
> Phone:(504) 988-7069 
> Fax:(504) 588-5326



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