DNA-Elution from gel
abrdlher at reading.ac.uk
Mon Jun 24 16:13:55 EST 1996
On Fri, 21 Jun 1996, Oliver Berkowitz wrote:
> Hi everybody,
> I´m searching for a fast and easy way of getting my DNA-fragments (about
> 1 kb) out of agarose-gels. At present I cut the bands out and spin the
> agarose-block on a Whatman-filter in a Eppendorf-tube. The effeciency is
> very poor. Is there anybody with a good technique ???
> Thanks for helping me, bye.
For that purpose, I did in the past some "electroelution".
I used short lengths of dialisis tubing, which had been previosly boiled
for 10 min in a solution of 2% NaHCO3 and 1mM EDTA pH8, rinsed in
distilled water, and boiled 10 min in 1mM EDTA pH8. Tubes were kept in
this solution at 4 degrees until whenever I needed them.
I put the slice of agarose in a piece of tube, closed by plastic clips,
in about 500ul 0.1xTBE buffer. Then I put the tube in a
minielectrophoresis tank, and run at 200V for 15 minutes, reversing the
polarity twice at the end for about 30 sec each time.
Then I pipetted out the buffer and precipitate the DNA with ethanol as
usual. The system worked ok for me.
Hope this helps
Nacho de las Heras
University of Reading
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