DNA extraction from formalin-fixed brain tissue
spbrown at man.ac.uk
Thu Apr 18 05:11:07 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
We have had lots of problems with formalin fixed brain tissue which had
been stored long term. The best method that we have found is by doing
'standard' DNA extraction but then doing 2 sets of PCR's...firstly PCR
with primers which flank then 'normal' PCR fragment...then secondly using
your standard primer set as nested primers to PCR from an anliquot of the
first reaction. You have to be extremely careful with controling for
contamination as this technique is extremely sensitive.
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