Best way to get DNA from gel bands?

Michael Myers myersm at rockvax.rockefeller.edu
Tue Jul 25 12:47:50 EST 1995


In article <24JUL95.15201632.0065 at SEMOVM.SEMO.EDU>, Allen Gathman
<C741SCB at SEMOVM.SEMO.EDU> wrote:

stuff deleted,
> 
> I know there are numerous kits, spin columns, etc. for this purpose.
> What is your experience with these?  What's the best (and reasonably
> cheap) method for getting DNA out of a band in a gel?
> 

This topic will probably go on for eternity. I know I've tried a zillion
approaches.

My current method: QIAquick from Qiagen. Faster than their Qiaex bead
method. Gel slice is melted in chaotropic salt (probably NaI), then spun
through a dna binding filter in a microfuge. Washed once, then eluted into
30 microliters of water or TE. It takes about 15 to 20 minutes, depending
on how big your gel slice is. Advantage here is that the eluted DNA is in
a small enough volume (and presumably free of EDTA) that you can proceed
with labeling or ligation rxns, no need to ethanol precipitate.

I have yet to test a new application of a Millipore product,the
Ultrafree-MC centrifugal filter device. You just spin the gel slice for 5
minutes, then ppt the eluate. Yields are not great for larger fragments,
but may be sufficient for most applications. The need to precipitate is a
pain. Check their ads in Science. You can call for a free sample (2 spin
units) at 1-800-MILLIPORE. Has anyone used it?

-- 
Michael Myers
The Laboratory of Genetics
The Rockefeller University
myersm at rockvax.rockefeller.edu



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