PCR from hair follicles/buccal(sp?) cells

Manuel Simon ma at ABC.univie.ac.at
Mon May 2 10:23:17 EST 1994


In article <01HBQPYDBQ3K8WWOD6 at MGRP.BGSM.WFU.EDU> BOWDEN at MGRP.BGSM.WFU.EDU writes:
>From: BOWDEN at MGRP.BGSM.WFU.EDU
>Subject: PCR from hair follicles/buccal(sp?) cells
>Date: 29 Apr 1994 06:09:08 -0700

>        We are going to try to carry out pcr analysis from samples sent by 
>mail. The idea is to test samples with 5-6 different microsatellite markers.
>For complicated reasons it will be difficult to get blood samples, but 
>hair follicles or buccal(sp.?) cell scrapings from the mouth are easy to 
>get. Can anyone suggest references for such a procedure, or relate there
>own experience? Do hair follicles, buccal smears travel well? That is could
>they be sent by regular mail? Also can you get reliable pcr from them?
>By the way, these are human samples. Thanks for any help which can be pro-
>vided.
>Don Bowden
>Dept. of Biochemistry
>Bowman Gray School of Medicine
>Winston-Salem, NC 27157

Hi, Don

We used to performe PCR from cells contained in the saliva for RFLP analysis 
some years ago. Probably there are better protocols availbale now. The 
protocol is very easy: your try to wash the cells (actually NOT really easy) 
in PBS. The cell pellet (from 2 ml saliva we obtained about 20 µl pellet) was 
then cooked with approx. 2 times (or more) of the pellet volume of essentially 
Promega 10x Taq-polymerase buffer (containing 1% Triton X-100)for e.g. 10 min. 
Then You set up your PCR by simply diluting this soupe 1:10. I cannot say 
whetherthe cells can stand US postal service. In addition, we finally stopped 
this procedure because of the high possibility of contaminations and 
uninterpretable results we got.

hope it helps
Manuel M. SIMON 



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