RNA from human skin

Gail Otulakowski gotulak at sickkids.on.ca
Wed Dec 9 11:52:09 EST 1998


Brunhilde Bloemeke wrote:
> 
> Has anybody a good protocol for isolation of total RNA from small human
> skin samples.  We tried many different methods but yields are very low.
> Thanks
The problem with trying to prep RNA from skin samples is that it is very
difficult to homogenize quickly and thoroughly.  We have had success
with the following trick:

Buy a stainless steel mortar and pestle (used normally for crushing rock
samples).  Chill it well in dry ice.  Freeze your skin sample in liquid
nitrogen.  Quickly transfer skin to mortar and pestle, and tap pestle
with hammer several times.  Remove pestle, break up clump of smashed
tissue quickly with a metal spatula, and hammer with pestle again.  This
process should yield a frozen powder.  Don't let it melt - transfer to a
guanidinium thiocyanate solution and homogenize with a small polytron. 
We have had success using the standard guanidinium/acid phenol method
(Chomczynski and Sacchi) but I suspect the commercial kits would also
function well, as long as the tissue is powdered first.  I have also
heard of people actually homogenizing tissue in liquid nitrogen with a
polytron (the liq N2 evaporates off at the end) but have never tried
this.

Gail Otulakowski,Ph.D.
Hospital for Sick Children
Toronto, Canada



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