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HELP: PCR and formaldehyde

Dirk dhunt02 at nospam.emory.edu
Fri Jan 7 10:48:55 EST 2000


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
>
>





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