I had formalin tissue chunks.
I first soaked the preservative out of the samples and digested the
sample down with Protease K.
First mince the tissue with a blade and soak for 24 hours in 1XTE
0.5% SDS (change this a couple of times during the 24 hours) The
sample should be on a rotator or rocker.
Then add protenase K to the tube every morning until the tissue
disolves. (Room Temp will work but I recomend at least 37° C or if
available 50°C)
>From there phenol extract as you normally do.
Then startedOn Fri, 03 Dec 1999 17:24:10 +0100, harvey
<harvey at ch1.fgg.eur.nl> wrote:
>It is no problem, just do it. We extract the DNA in tail mix buffer
>which is:
>>>50mM Tris pH8
>100mM EDTA
>100mM NaCl
>1% SDS
>>To 400ul of tail mix we add proteinase K
>with proteinase K (10-20ug) at 55 degree C overnight
>Phenol Cholorform and ethanol precipate
>>"Parent, Eric" wrote:
>>> Dear netters
>>>> I would like to know if anybody has experience doing PCR on samples
>> preserved in formaldehyde, if it can be done.
>>>> Any help would be greatly appreciated
>>>> Eric
>>>> Eric Parent
>> Maurice Lamontagne Institute
>> Fisheries and Oceans Canada
>> Invertebrates and Experimental Biologie
>> 850 route de la mer
>> Mon-Joli, Qc
>> Canada
>> G5H 3Z4
>>parente at dfo-mpo.gc.ca>>http://www.qc.dfo-mpo.gc.ca/iml/fr/intro.htm>>--
>-------------------------------------------------------
>K Harvey
>Dept of Cell Biology
>Erasmus University Rotterdam
>The Netherlands
>mailto:harvey at ch1.fgg.eur.nl>>
