IUBio

DNA isolation from agarose gels

Hiranya S. Roychowdhury hroychow at NMSU.EDU
Fri Mar 12 11:09:24 EST 1999


At 10:35 PM 3/11/99 GMT, jag100 at snethen.com wrote:


>I use AgarAce from Promega to extract large (5-10 kb) PCR fragments from
>agarose.  This works alright, but could be better.  I would like to try some
>of the protocols suggested in this thread, but I am doing this in high
>quantity (upwards of 70-80 fragments at a time)--it would be a bit daunting
>to prepare individual tubes each time (I am in a 96-well block now.)  Any
>ideas for high- throughput (and still high-yield) isolations?
>
>Thanks,
>
>Jen
>jag100 at snethen.com
>
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>
I am certain that you can adapt the "freeze squeeze" method that others and
I posted earlier for a 96-w plate, given you have the necessary centrifuge
that handles the plate. However, even if you purchased the spin columns
(available from regular suppliers, I think) you'll be saving a lot of money
while obtaining great yield. Also, check out D.Kim's post where he talks
about essentially the same thing as I do with my home-made microfuge
columns, but with cheap, non-hydrophobic, barrier tips. You don't have to
make the columns. Just put your gel slice in the wide end of the tip and use
it as the column. BUT, you slices have to be small.

Hiranya.


Dr. Hiranya Sankar Roychowdhury
GENE LAB/ EPPWS
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu




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