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Updated: Jun 3, 2026

In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions
Published on: July 17, 2019
Detection of DNA sequences using biotinylated probes
J L Woodhead1, H Figueiredo, A D Malcolm
1Department of Biochemistry, Charing Cross and Westminster Medical School, London, UK.
Abstract:
It was found (1) that biotinylated analogs of dUTP, which contain a biotin molecule covalently bound to the C-5 position of the pyrimidine via an allylamine linker arm, can be used as substrates for DNA polymerase. Figure 1 shows the structure of biotin-11-dUTP, a commonly used substrate for biotinylation by the method of nick translation (2). The substituted DNA molecule will have similar hybridization properties to the un-substituted DNA. After hybridization to the test DNA, the biotin probe can be located by a variety of methods, including incubation with streptavidin-peroxidase conjugate, which will be described here. Fig. 1. Structure of biotin-11-UTP.
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