Bernd Dr. Reckmann

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.

Quantitative Analysis of Polymerase Chain Reaction (PCR) Products by Means of Fluorescence Detection

PCR primers covalently labeled with biotin and a fluorescent dye allow immobilization and separation of the products which, subsequently, can be quantitatively analysed. 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 and with internal standardization. PCR is carried out in a newly developed thermocycler with exceptional heating and cooling rates. The thermocycler performs 30 cycles in 2 hours.