DNA from Polyacrylamide & Protein Fusion and Purification System

Basavaraju Shankarappa bsh at MED.PITT.EDU
Wed Dec 9 18:53:01 EST 1992


>   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




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