Real Time PCR Using Taqman Probe | RT PCR using Taqman Dye |

An oligonucleotide probe called taqman probe is constructed containing a fluorescent reporter dye on the 5' end and a quencher dye on the 3' end. While the probe is intact, the proximity of the quencher dye greatly reduces the fluorescence emitted by the reporter dye by FRET.

If the target sequence is present, the probe anneals downstream from one of the primer sites and is cleaved by the 5' nuclease activity of Taq DNA polymerase as this primer is extended.

This cleavage of the probe:
-- Separates the reporter dye from the quencher dye, increasing the reporter dye signal.
-- Removes the probe from the target strand, allowing primer extension to continue to the end of the template strand. Thus, inclusion of the probe does not inhibit the overall PCR process.