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Nucleic Acids Research, 1989, Vol. 17, No. 12 4605-4610
© 1989

CHEMISTRY

Labelling oligonucleotides to high specific activity (I)

Alister G. Craig*, Dean Nizetic and Hans Lehrach

Genome Analysis Laboratory, Imperial Cancer Research Fund Laboratories Lincoln's Inn Fields, PO Box 123, London WC2A 3PX. UK

Received March 8, 1989. Revised May 16, 1989. Accepted May 16, 1989.

The normal procedure for labelling oligonucleotides radioactively is the use of polynucleotide kinase and {gamma}32P–ATP. However, this has the disadvantage of only introducing one labelled base per molecule of the oligonucleotide. In this paper we describe an approach based on primer/template combinations using conventional fill–in conditions followed by the release of the labelled sequence by digestion with uracil-DNA glycosylase.

*Present address: Molecular Parasitology group, University of Oxford, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK


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