DNA from hair

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Thu Apr 30 12:25:20 EST 1992


In article <9204282213.AA28462 at trog.biol.sfu.ca>, cmuir at TROG.BIOL.SFU.CA (C. Cam Muir) writes:
> I writing to see if anyone out there has experience
> extracting DNA from hair. I am having troubles with it.
> I want to go directly to PCR with the extract. Thanx, in
> advance for the help.
> 
Hi!  I have tried it and I had no trouble getting the DNA.  I DID have a
problem with my primers being contaminated with cloned cDNA, but that is
another story.  Anyway, there was a very good protocol for extraction of DNA
from very small samples (such as a single hair follicle) in Technique 1:96-102
by B. Hopkins, J.E.N. Morten, J.C. Smith, and A.F. Markham entitled "The
development of methods for the analysis of DNA extracted from forensic
samples".  They prepared enough DNA from these small samples to do single locus
probe analysis via Southern blotting and hybridization.  I could get a copy of
the paper to you so that you could see how great their results were, but here
is a direct reproduction of one of their protocols:
Protocol 5
DNA extraction from hair roots
1.  Suspend hair roots (up to 50) in 500 ul of 10mM Tris HC., pH 8.0, 100mM
NaCl, 1mM EDTA (STE).
Add 25 ul 10% SDS.
Add 20 ul Proteinase K (10 mg/ml).
Incubate for 2 h at 56C.
2.  Add 0.1 volume 2M sodium acetate, pH 5.6.
Extract with phenol/chloroform (2X).
Extract with chloroform (1X).
Add 10 ul glycogen (10 mg/ml) to the aqueous phase.
Add 1 ml ethanol.
Mix by gentle inversion.
3.  Centrifuge for 10 s.
Decant supernatant.
Resuspend pellet in 0.2M sodium acetate pH 7.0 (200 ul).
Add 500 ul ethanol.
Mix by gentle inversion.
4.  Centrifuge for 10 s.
Decant supernatant.
Add 1 ml 80% ethanol.
Centrifuge for 10 s.
Decant supernatant.
5.  Dry pellet under vacuum for 5 min.
Dissolve pellet in appropriate restriction enzyme buffer.

At step 5, I just dissolved my pellet in pure dH2O and went on to PCR.  This
paper also has protocols for blood, semen, urine, saliva, mixed blood & semen,
and stained material.  They also did a few studies of storage of the samples
under various conditions before attempting to make DNA to try to simulate a
crime scene.
Hope this helps! 

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Barbara Bugg, M.S.  bugg at mbcf.stjude.org       "I refuse to have a battle of
St. Jude Children's Research Hospital          wits with an unarmed person."
Memphis,  TN  USA
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