Nick Translation

Abstract

Nick translation is a rapid and easy method of labelling duplex DNA with radioactive or chemically modified nucleotides. The technique is based on the ability of DNase I to introduce random nicks in the target DNA and the simultaneous exonuclease and polymerase activity of Escherichia coli DNA polymerase I to replace existing nucleotides with labelled nucleotides (Kelly et al., 1970; Rigby et al., 1977).

Keywords: DNA labelling; DNase I; nick translation

Figure 1.

Labelling of duplex DNA by nick translation: (a) reaction at 16°C; (b) reaction at 20°C or above.

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References

Kelly RB, Cozzarelli NR, Deutscher MP, Lehman IR and Kornberg A (1970) Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. Journal of Biological Chemistry 245: 39–45.

Rigby PWJ, Dieckmann M, Rhodes C and Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113: 237–251.

Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

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How to Cite close
Woodward, K(Jul 2003) Nick Translation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003748]