DNA extraction from formalin-fixed brain tissue

Pamela J Snyderer snyder.9 at postbox.acs.ohio-state.edu
Sun Apr 14 21:23:03 EST 1996


In article <3173992B.6ED2 at shef.ac.uk>, "P.Orange" <P.Orange at shef.ac.uk> wrote:

> Hello, A collegue and I have been trying for ages to try and extract DNA 
> from long term storage formalin fixed brain tissue.  We have tried 
> standard Phenol/Chloroform techniques (including greatly extended 
> proteinase treatment), DNAzol, Puregene kit, all to no avail.  All we 
> need is enough DNA to do PCR for a 250bp fragment.
> Does anybody therefore hold the holy grail of DNA extraction, we are 
> willing to give any kit or technique a go.
> Please help us,
> Paul Orange & Claire Shepherd

Are you rehydrating the tissue before you try to get into it?  I routinely
do PCR from formalin fixed tissue, paraffin embedded tissue and even
paraffin embedded tissue from slides.  

I find it helpful to work with small amounts of tissue (10 - 30 mg)

I start with 100% ethanol x 2, then 80%, then 50%, then ddH2O

from there proteinase K and SDS usually gets me in, Homogenization is
usually not necessary esp. if tissue is thinly sliced or minced.

I have found that tempermental primer sets sometimes will not tolerate
this kind of (generally) degraded templates.



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