Extracting DNA from low-melt agarose gels

[Sebastian Bunka]

> Date:          8 Jul 93 12:03:12 GMT
> To:            "bionet.molbio.methds-reagnts mail newsgroup" <bionet-news at daresbury.ac.uk>
> From:          KARN at butler.edu
> Reply-to:      KARN at butler.edu
> Subject:       Extracting DNA from low-melt agarose gels

> I wish to use low-melt agarose gels to clean up a PCR product of about 1 kb
> (+/- 0.2 kb) but would like to avoid the multiple extraction procedure
> described in Maniatis.  My goal is to sequence the product so it must be
> relatively clean.  Anybody got an improvement on the Maniatis technique
> or must I haul out the phenol?
> Bob Karn
> KARN at BUTLERU.edu
>
>
>
A very good method for rapid and high purity recovery from standard
agarose gels is the Geneclean II Kit from BIO 101 (LaJolla, CA, Ph.
(800) 424-6101 or (the same) from USB "USBioClean".
Both Kits work with TBE or TAE buffered gels. Short description:
run gel, cut out desired band, add NaI (+TBE modifier) heat 5 min
50 C, add Glassmilk, store on ice 5 min, wash 3x w/ washbuffer, add
a. dest., incubate 5 min., spin down, recover pure DNA.

I have used both kits, and they were very useful for repeated RE
digests, ligation reactions and the recovery % is about 80%!
Kits cost about 100-120 US$ and allow about 200-250 purifications

have fun
Sebastian
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|Dr. Sebastian Bunka      Email: Sebastian.Bunka at vu-wien.ac.at     |
|Inst.f.Bakteriologie                                              |
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|A-1030 Wien                                                       |
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