=?iso-8859-1?Q?R=E9f=2E_=3A_Re=3A_RTPCR_from_paraffin_sections-_still?= looking for that somebody!!
I'm a new subscriber and that was my first question, nice to have some
quick responses. Thanks Nina and Bob but I don't believe mRNA degradation
can explain our whole problem. Firstly (I didn't include this info in my
initial messsage) In situ hybridization with riboprobes works even better
in our paraffin sections (5µ) than our frozen sections (15µ), so in
paraffin sections we have 3 times less material but twice the signal. Its
true that PCR requires intact mRNA while ISH doesn't. Secondly we are also
unable to amplify genomic DNA from our paraffin sections. So we think its
more a question of digestion/extraction/accessibility of mRNA molecules.
We're now looking and testing extraction protocoles. We believe RTPCR from
paraffin sections is possible because the only three makers of
microdissection systems : LEICA, ARCTURUS, and PALM claim that it's
feasable. And thats why we bought the dam- !$#O*!?X ! $# machine.
Somebody, Somebody out there must know how to do it.....
Molecular Biology Scientist
B.P. 37 Labége Innopole
31676 LABEGE CEDEX FRANCE
tel : (33)561004179 fax :(33)561004001
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