If the tissue is not embedded, but straight out of the fixative, wash the
chopped up sample for a couple of hours in several changes of sterile water
and then proceed as usual to the digestion step (spin down before each
change to remove as much of the water as possible).
Extraction of DNA for PCR from long stored formalin fixed, paraffin wax
embedded tissues can be a problem, as the DNA becomes degraded over time,
(I tried to get PCRable DNA from blocks embedded in 1942, with no joy) and
standard phenol/coloform extraction (ref Maniatis) might be better and give
larger yield than the Quigen kit.
Hope this helps
>From: Dolors Fuster
>Date: Mon, 22 Jul 2002 20:26:41 +0100
>Hi histonetters !
>What do you recommend us to obtain the best results in DNA extraction
>from formalin fixed tissue? should we do any kind of treatment before
>extraction? (Water rinses, alcohol changes....)
>We are usin Qiagen kit with good results for parafin embeded samples
>fresh tissue and other kind of samples (cell culture, serum...). but
>are not successful with formalin fixed tissue
>Thanks in advance
>Tecnic Especialista en
>Departament d'Anatomia i Embriologia Humana
>Facultat de Medicina (HCP)
>Universitat de Barcelona
Bone Research Unit
Tel & fax +27 11 717 2298
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