Labelling DNA in vitro — Nick Translation
If a single-stranded DNA molecule is placed in contact with a complementary single-stranded DNA sequence, the two molecules will associate with one another by hydrogen bonding between the bases on their respective strands. This association, or hybridisation, forms the basis of very powerful techniques for detecting and quantifying specific nucleic acid sequences. Whether the hybridisation is done in solution or to nucleic acid immobilised on filters, a radioactively labelled probe will be required. The enzyme DNA polymerase I from E. coli catalyses a reaction which can be used to replace existing unlabelled nucleotides in DNA with radioactive ones. The reaction has been called nick translation, and is very widely used in molecular biology.1,2 It has nothing whatever to do with the translation of messenger RNA into a polypeptide chain on the ribosomes.
KeywordsHigh Specific Activity Nick Translation Active Chromatin Tissue Culture System Label Nucleotide
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© John M. Walker and Wim Gaastra 1983