RT-PCR on formalin fixed paraffin sections

Alex Dobrovic adobrovic at medicine.adelaide.edu.au
Mon Mar 24 01:01:03 EST 1997


I would appreciate input from anyone who has experience with isolating RNA
from formalin fixed paraffin embedded sections and performing RT-PCR on the
RNA.

The sections we want to use have been stained in haematoxylin and eosin,
will this inhibit RT/PCR or both ? The PCR product is approximately 230 bp
and we need to get results using 5um sections.

I am after advice of any kind e.g. -

(1) Removing the parrafin
(2) Carrier for precipitating RNA
(3) RT enzyme
(4) specific or random primed RT
(5) how many rounds of PCR

Thanks in advance,

Damian Hussey BSc (Hons)



Alexander Dobrovic, Ph. D.
Chief Scientist
Department of Haematology-Oncology
The Queen Elizabeth Hospital
Woodville, SA 5011, Australia

Affiliate Senior Lecturer
Department of Medicine
University of Adelaide

Tel 61-8-8222 6884
Fax 61-8-8222 6046 (note new numbers as of August 1996)





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