DNA from faeces
Andre Hamel
hamel at cc.umanitoba.ca
Sat Jan 21 12:13:34 EST 1995
I recommend homogenizing feces in 6 M GITC on ice -> centrifuge
to pellet out debris -> transfer supernatant to new tube & centrifuge
again (10,000 xg for 10 min at +4 to +10*C), followed by adding a slurry of
silica/diatomaceous earth to the supernatant in order to bind the DNA,
followed by washing away the "crap" (pun intended) with 50% ethanol/TE,
then elute DNA with TE ... can use right away or precipitate with 0.3 M
NaAc and 2x vol ethanol.
Several rather decent versions have been posted to this newsgroup (in
the FAQ? ... otherwise check the bionet archives). I like using spin
cartridges (small units that fit into top of 1.5 mL microfuge tube ...
have porous mesh/frit that catches the glass particles ... allows for
thorough washing.
These centrifuge tube frit/mesh units are available from several
manufacturers for low cost AND are reuseable (wash by first flushing in
warm water, then drying with 100% alcohol (your choice ... methanol,
ethanol, propanols) ... air dry then flush & soak for few minutes in
chromic-sulfuric acid then rinse well with water (chromic-sulfuric acid = 25
mL of 50% w/v aqueous solution of chromium trioxide slowly stirred into
4 L of conc sulfuric acid ... this chromic-sulfuric acid lasts for a
long time if NOT reused once wetted with water (we keep reused stuff in
separate bottle ... once it becomes green in color it is no longer
useable AND is more toxic).
I've recently converted to this method for getting PCR quality DNA from
animal feces (herbivores) compared to standard GITC/phenol/CHCl3 ext'n
(which doesn't remove PCR inhibiting compounds).
I wish to once again thank those who suggested my using this method for
feces DNA.
cheers,
********************
Andre Hamel email: hamel at cc.umanitoba.ca
Manitoba Government tel: 204/945-7630
Department of Agriculture FAX: 204/945-8062
Veterinary Services Branch
Winnipeg, Manitoba, CANADA
********************
In <3fon5s$s58 at infoserv.rug.ac.be> dieter.deforce at rug.ac.be writes:
>I would like to know if it is possible to extract human genomic DNA from
> faeces. Can anybody tell me where I can find a protocol or an article
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