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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