DNA elution from PAGE
James Zacka
jzacka at worthington-biochem.com
Tue Jun 10 08:01:42 EST 1997
Worthington Biochemical, www.worthington-biochem.com, has a new, rapid
Gel-Spin DNA Recovery kit which works well for both DNA or RNA recovery.
Jim Zacka/Worthington
In article <01bc7595$0ae3a800$467df88f at KAISTWF.kaist.ac.kr>, "Yeon Chul
Kim" <e_yckim at cais.kaist.ac.kr> wrote:
> Thilo Hofmann <101575.1370 at CompuServe.COM>ÀÌ(°¡)
> <5n0gpb$9va$1 at mhafc.production.compuserve.com> ±â»ç¿¡¼ ÀÛ¼ºÇß½À´Ï´Ù...
> > Hi!
> >
> > I´m looking for a protocol to elute a DNA band out of a
> > Acylamide Gel stained with Ethidium bromide.
> > I will cut out a particular band and then elute the
> > DNA in TE or some other buffer.
> > The DNA size is something around 400 - 500 bp
> >
> > Bernd Eschweiler
> > Institut für Wasserforschung
> > Germany
> > ifw at poboxes.com
> >
> > --
> > Thilo Hofmann
> > Zum Kellerbach 56
> > 58239 Schwerte
> > email: ifw_mail at compuserve.com
> >
> >
>
> The protocol is in "Molecular Cloning:A lab. manual" edited by Sambrook et
> al.
>
> 6-24~6-27.
>
> Breif description of my modified method
>
> 1. Cut DNA band
>
> 2. Make 2% agarose gel and make an incision in the gel
>
> 3. Cut the ion exchange paper (DE81, Whatman Cat#3658 023) slighty wider
> than band.
>
> 4. Attach band onto paper.
>
> 5. Perform electrophoresis for ca 10 min.
>
> 6. Check paper is red (using UV)
>
> 7. Transfer paper into EP tube which have small hole at bottom
> (small hole was made using needle)
>
> 8. Wash paper with TE
>
> 9. Elute DNA with TE + 5M Nacl 150 ul (3 times, so total is 450 ul)
>
> 10. phenol/CHCl3 extraction
>
> 11. EtOH down.
>
> This protocol give good results for various DNA size.
>
> In my case I purified 30 bp DNA.
>
> Good-Luck..
>
> Yeon Chul Kim
More information about the Methods
mailing list