The best purified fragment I have beem obtained don't use any of
this kind of kit. I use the following procedure:
1) - Run an acrylamide gel. Concentration based on the size of
2) - Passive elution over nigth in 1x TE, with constant
agitation. Use 200 to 500 microliter.
3) - Dry out the TE in a speed vac.
4) - Resuspend the DNA fragment in 15 microliter of 1x TE.
5) - Use this DNA for sequence.
This procedure give me very clean DNA band.
On 2 Dec 1995, Eric C. Anderson wrote:
> In article <Pine.A184.108.40.2061201072050.45807A-100000 at umabnet.ab.umd.edu>,
> "Mary P. Remington" <mremingt at umabnet.ab.umd.edu> wrote:
>> > I would like some input on the best methods for DNA fragment
> > purification methods. I am currently using Qiaex but, am wonder about
> > the spin column methods. Thanks, Mary
>> if you want to spend the money (which you apparently do if you're already
> using Qiaex), try the Qiaquick columns from Qiagen. they are about 3X as
> fast as Qiaex II (or Geneclean) and the purity is similar with a slightly
> lower yield than Qiaex.
>> i have no connections to qiagen other than as a customer.
> Eric C. Anderson
> Cell Biology and Genetics
> Memorial Sloan-Kettering Cancer Center
> Sloan-Kettering Institute
> 1275 York Ave., Box 470
> New York, NY 10028
> Phone: (212)639-2977
> Fax: (212)717-3298
>e-anderson at ski.mskcc.org>>