Efficient purification of DNA fragments using a protein binding membrane
Author(s) -
Øistein Ihle
Publication year - 2000
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/28.16.e76
Subject(s) - biology , dna , microbiology and biotechnology , endonuclease , hapten , restriction enzyme , biochemistry , antibody , genetics
A novel and efficient method has been developed for isolation of correctly digested DNA fragments without the use of classic size-dependent electro- phoretic separation methods. To achieve this, DNA fragments are end-labelled by haptens. After specific endonuclease digestion of the hapten-labelled DNA, the DNA is incubated with a protein that specifically binds to the hapten. The incubation mixture is then passed through a cartridge containing a protein- binding membrane that does not bind DNA. Undigested and partly digested DNA are retained on the membrane, while correctly digested DNA is selectively recovered for use in further downstream applications.
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