Abstract
Nick translation was the first method devised for the in vitro labeling of DNA (1). During the reaction the DNA to be labeled is nicked by DNase I yielding a free 3′ hydroxyl end. DNA polymerase I then adds a new nucleotide to this end. The 5′–3′ exonuclease activity of the polymerase then moves the “nick” along the strand in the 3′ direction. The addition of a radioactively labeled nucleotide to the reaction results in probes that can be used in hybridization reactions to DNA immobilized on Southern blots, colony lifts, etc.
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References
Kelly, R. B., Cozzarelli, N. R., Deutscher, M. P., Lehman, I. R., and Kornberg, A. (1970) Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. J. Biol. Chem. 245, 39–43.
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© 2002 Humana Press Inc.
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Reid, A. (2002). Nick Translation. In: de Muro, M.A., Rapley, R. (eds) Gene Probes. Methods in Molecular Biology, vol 179. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-238-4:023
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DOI: https://doi.org/10.1385/1-59259-238-4:023
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-885-1
Online ISBN: 978-1-59259-238-8
eBook Packages: Springer Protocols