Efficient purification of DNA fragments using a protein binding membrane.
A novel and efficient method has been developed for isolation of correctly digested DNA fragments without the use of classic size-dependent electrophoretic 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.
Ihle
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National Institute of Public Health, Department of VAIM, PO Box 4404 Torshov, 0462 Oslo, Norway. oistein.ihle@folkehelsa.no
Michaelsen
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D001710:Biotin; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli; D019793:Fluorescein; D006241:Haptens; D008567:Membranes, Artificial; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011485:Protein Binding
D004268:DNA-Binding Proteins; D006241:Haptens; D008567:Membranes, Artificial; D001710:Biotin; D004247:DNA; D019793:Fluorescein