DNA from Polyacrylamide & Protein Fusion and Purification System

Victor Jongeneel vjongene at isrec-sun1.unil.ch
Thu Dec 17 02:12:00 EST 1992

Basavaraju Shankarappa (bsh at MED.PITT.EDU) wrote:
: >   Hi! I was wondering if anyone out there knows of any company that sells a kit
: > for extracting DNA from polyacrylamide gels. According to Molecular Cloning
: > (2nd Edition) by Sambrook et al on page 6.46, there exists a "Crush and Soak"
: > method. However, the method is lengthy and inefficient. With so many kits
: > available for extracting DNA out of agarose gels, I haven't been able to find
: > one that's for use with polyacrylamide gels. Unfortunately, I must run my DNA
: > samples on PAGE. I would appreciate it if anyone could provide any info. or
: > even suggest a better method other than "Cursh and Soak".
: > Michael Cheng
: > Dept. of Biological Chemistry
: > Kyoto University
: > a52041 at sakura.kudpc.kyoto-u.ac.jp

: There is a system of solubilizable acrylamide gels for isolation of DNA
: fragments by Dr. Norman Hansen.  He was our teacher and he was a great
: one.
: 	In this system, the gels are crosslinked by disulfide bridges and
: the gels are supposed to be quite effective in resolving MW ranges from
: 21 kb down.  I do not know whether anybody markets this system.  You can
: obtain the disulfide containing bis from Bio-Rad.  The gels are solubilized
: by treating with mercaptoethanol and the DNA purified by DEAE cellulose. 
: I guess you should be able to clean DNA with other methods also.
: 	The reference is J.N. Hansen.  Use of solubilizable acrylamide 
: disulfide gels for isolation of DNA fragments suitable for sequence
: analysis.  Anal. Biochem. 116: 146-151 (1981).

: Raj Shankarappa
: bsh at med.pitt.edu


Another (and easier) way is to electro-elute your DNA from the gel.
There are several commercial systems available to do this.  We have
tried the BioTrap from Schleicher & Schuell (may be sold under
different trade names outside Europe) and a "salt trap" model sold by
IBI/Kodak.  They both work fine, but I prefer the IBI system, which is
quicker and allows you to elute up to 6 samples simultaneously.  All
you have to do after elution is EtOH precipitation.

Hope this helps.  Good luck!


C. Victor JONGENEEL                       vjongene at isrecmail.unil.ch
Ludwig Institute for Cancer Research, Lausanne Branch
Chemin des Boveresses 155                       FAX: +41-21-653-4474
CH-1066 EPALINGES				Tel: +41-21-653-6275

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