Quantitative analysis of polymerase chain reaction (PCR) products using primers labeled with biotin and a fluorescent dye.
PCR primers covalently labeled with biotin and a fluorescent dye allow immobilization and separation of the products which can be quantitatively analyzed subsequently. The procedure we have developed circumvents electrophoretic separation and radioactive labeling. Exact quantitative analysis of reaction products is feasible during the logarithmic phase of amplification when Taq polymerase is not limiting, as it is during the plateau phase of the reaction. With appropriate standardization the procedure can be used for routine diagnostic purposes.
Landgraf
A
A
Abt. Biophysikalische Chemie, Zentrum Biochemie, Medizinische Hochschule Hannover, Germany.
Reckmann
B
B
Pingoud
A
A
D001483:Base Sequence; D001710:Biotin; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D005091:Exons; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic; D013861:Thiocyanates
D004277:DNA, Single-Stranded; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013861:Thiocyanates; D001710:Biotin; D004247:DNA; D016650:Fluorescein-5-isothiocyanate