formalinfixed-PCR

Zbigniew Rudzki mprudzki at cyf-kr.edu.pl
Fri Aug 7 07:46:09 EST 1998


Marianne Schwartz wrote in message <35CA2B2B.A03E4ABF at post6.tele.dk>...
>I am in the need for a reliable method to do PCR on a
>formalinfixed-paraffin embedde liver sample. Can anybody help me with
>this. PCR product app. 150 bp.


Dear Marianne,

this is currently also my problem.  I am following roughly the protocols of
Diaz-Cano et al. (Diagn Mol Pathol 6: 342-6, 1997) and Isola and al. (Am J
Pathol 145:1301-8, 1994).  They have been optimized for DNA extraction from
archival paraffin-embedded material.
So far I have got 150-250 microgram of DNA/sample (twenty 5-micron thick
sections from each block) of the average length around 450-500 bp.  My
oldest blocks are from 1975 and most of them had been overfixed (several
days in formaline).  I work on large bowel but the actual material should
not matter a lot.
The next week I will fight with PCR -- if you are interested in more close
details of extraction and PCR outcomes [if the are any ...] pleas let me
know by e-mail.

I hope this may help

Zbigniew Rudzki
Dept. Pathol., Jagiellonian University,
Krakow, POLAND





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