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• W2030204860 endingPage "928" @default.
• W2030204860 startingPage "917" @default.
• W2030204860 abstract "A novel highly sensitive non-radioactive DNA labeling and detection system based on the ELISA principle has been developed. DNA is modified with the cardenolide-hapten digoxigenin by enzymatic incorporation of digoxigenin-labeled deoxyuridine-triphosphate with Klenow enzyme. Digoxigenin is linked to dUTP via an 11-atom linear spacer (Dig-[11]-dUTP). Following hybridization of membrane-bound target-DNA with a digoxigenin-labeled probe, the hybrids are detected by an ELISA reaction using digoxigenin-specific antibodies covalently coupled to the marker enzyme alkaline phosphatase [(Dig):CIAP]. This binding of antibody: marker enzyme-conjugate is followed by an enzyme-catalysed coupled redox reaction with the colour substrates 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitroblue tetrazolium salt (NBT) giving rise to a deep-blue coloured, water-insoluble precipitate directly adhering to the membrane. The digoxigenin system allows the detection of 0.1 pg homologous DNA within 16 h in dot- and Southern-blots on nitrocellulose or nylon membranes avoiding any significant background even after a prolonged period of color development. Due to its high sensitivity and specificity, the new system is appropriate for detection of single-copy genes in genomic blots as well as for Northern, slot, colony, plaque and in situ hybridizations." @default.
• W2030204860 created "2016-06-24" @default.
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• W2030204860 date "1990-01-01" @default.
• W2030204860 modified "2023-09-27" @default.
• W2030204860 title "Non-radioactive Labeling and Detection of Nucleic Acids. I. A Novel DNA Labeling and Detection System Based on Digoxigenin: Anti-Digoxigenin ELISA Principle (Digoxigenin System)" @default.
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• W2030204860 doi "https://doi.org/10.1515/bchm3.1990.371.2.917" @default.
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