Best DNA fragment purification methods.

Luiz Carvalho carvalho at BIODEC.WUSTL.EDU
Sat Dec 2 20:47:54 EST 1995


Hi,

	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 
your fragment.
	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.

	Good luck.

	Luiz

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On 2 Dec 1995, Eric C. Anderson wrote:

> In article <Pine.A32.3.91.951201072050.45807A-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
> 
> -- 
> 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
> 
> 



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