Quantitative measurement of nonisotopically labeled polymerase chain reaction product.
A novel approach for evaluation of polymerase chain reaction product was described. A biotin-labeled DNA (or RNA) probe is hybridized to the digoxigenin-labeled target PCR product. The hybrids are retained on an anti-biotin antibody-coated plate. An anti-digoxigenin Fab conjugated with alkaline phosphatase is used for generating either a fluorescence or a chromagenic substance. Both fluorogenic and chromagenic systems can be used for quantitative measurement of the PCR product/probe hybrids. The method is specific, fast, nonisotopic, sensitive, quantitative, and practical.
Yang
B
B
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Viscidi
R
R
Yolken
R
R
D001483:Base Sequence; D001710:Biotin; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes; D004076:Digoxigenin; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015347:RNA Probes; D012680:Sensitivity and Specificity
D015342:DNA Probes; D015347:RNA Probes; D001710:Biotin; D004247:DNA; D004076:Digoxigenin