DNA-Extraction

Caron Sebastien carons at MAGELLAN.UMONTREAL.CA
Mon Sep 20 10:13:41 EST 1999



On Fri, 17 Sep 1999, Silke Stoll wrote:

> Hallo to everybody,
> I am looking for a method to extract the DNA from a Polyacrylamid-Gele.
> I am using this kind of geles, beause I am working with the Differential
> Display.
> Thank you very much,
> Silke Stoll
> 
> mailto:sastoll at uni-muenster.de
> 
> 
> 

Here's what we do in the lab. With a new scalpel blade, cut out the gel band of
interest and put in a tube with a screw cap. Add 100 ul deionized water and
incubate 1h in a water bath at 70 Celcius, then leave overnight on the bench.
Quickspin and transfer the water to a new eppendorf tube. Precipitate 1h at
-20 with 5 ul glycogen (10 mg/ml), 10 ul NaOAc 3M ph 5,3 and 450 ul ETOH
100%. After centrifugation, wash the pellet with 85% ETOH instead of
70% because if the % of ethanol is too low, your small DNA fragments
will disolve themselves in it. Resuspend in 12 ul water. Use 4 ul for
reamplification PCR.

Sebastien Caron
Institut de Recherche en biologie vegetale
4101 Sherbrooke est,
Montreal, Quebec
H4C 1Y1




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