DNA-Elution from gel
yoonpark at u.washington.edu
Fri Jun 21 18:12:33 EST 1996
I guess you don't want to buy commercialized kits such as GeneClean from
Bio101. If it's the case, here is a simple protocol you can try.
1) Cut the band from agarose gel. Blot excess buffer from the surface of
the gel fragment and place it in a microfuge tube. Cut the gel
frgment into small piece and distribute to more than one microfuge
tubes if necessary (ca. 300ul of gel/tube).
2) Add 200ul of phenol, and vortex.
3) Freze the sample in a dry ice-ethanol bath for 5 min.
4) Centrifuge for 15 min to pellet the agarose.
5) Remove the top aqueous layer. This is the extuded buffer which
contains the recovered DNA.
6) Add 200ul of TE to Agarose-henol remaining in tube.
7) Vortex to reextrac and repeat step 5-6.
8) Phenol-extract the agueous layer twice.
9) EtOH precipitation of the recovered DNA.
Ref.) T.J. Silhavy, M.L. Berman, and L.W. Enquist (1983) Experiments with
Gene Fusion. Cold Spring Harbor Laboratory.
University of Washington
Dept. of Oral Biology, Box 357132
Seattle, WA, 98195-7132
E-Mail: yoonpark at u.washignton.edu
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.
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