Detection of DNA Sequences Using Biotinylated Probes
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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.
