title: Highly sensitive biotin-labelled hybridization probe.
abstract: A simple chemical method for introducing biotin into nucleic acids has been developed for the synthesis of nonisotopic hybridization probes. The method is based on the reaction of biotin hydrazide with amino residues of nucleic acids by using glutaraldehyde as a bifunctional coupling reagent. Biotin-labelled deoxyribonucleic acid (DNA) was detected by the use of alkaline phosphatase-labelled avidin, and alkaline phosphatase activity was measured by colorimetric and chemiluminescence methods. The chemiluminescence method using the nicotinamide adenine dinucleotide phosphate (NADP)/alcohol/alcohol dehydrogenase/microperoxidase/isoluminol system gave the highest sensitivity. A few picograms of lambda-phage DNA coated on a microtiter plate well could be detected by this method.
authors: H
authors: H
authors: Arakawa
authors: M
authors: M
authors: Maeda
authors: A
authors: A
authors: Tsuji
authors: T
authors: T
authors: Takahashi
mesh_terms: D001710:Biotin; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D007202:Indicators and Reagents; D009693:Nucleic Acid Hybridization
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D007202:Indicators and Reagents; D001710:Biotin