TaqMan (RT-PCR, real time PCR)

A fluorogenic 5′-nuclease assay using a FRET probe, generally consisting of a green fluorescent dye at the 5′-end and an orange quencher dye at the 3′-end of a DNA. In a PCR process when the probe anneals to the complementary strand, the Taq polymerase cleaves the probe and the dye molecules are separated. Thus, the quencher can no longer suppress the reporter (e.g., the green dye) and a fluorescence detector can quantitate the green emission and the green fluorescence directly correlates with the yield of the PCR product. In seven minutes, sufficient quantities of DNA can be produced for the identification of pathogenic microbes. This technique may be used also for the molecular definition of deletions and SNIPs. Taq DNA polymerase, FRET, PCR, quenching, SNIPS; Medhurst AD et al 2001 Brain Res Mol Brain Res 90(2):125; Ranade K et al 2001 Genome Res 11:1262.