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added to the PCR reaction mixture. This probe is an oligonucleotide with a reporter dye attached to the 5'
end and a quencher dye attached to the 3' end. Till the time the probe is not hydrolized, the quencher and
the fluorophore remain in proximity to each other, separated only by the length of the probe. This proximity
however, does not completely quench the flourescence of the reporter dye and a background flourescence is
observed. During PCR, the probe anneals specifically between the forward and reverse primer to an internal
region of the PCR product. The polymerase then carries out the extension of the primer and replicates the
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template to which the TaqMan® is bound. The 5' exonuclease activity of the polymerase cleaves the probe,
releasing the reporter molecule away from the close vicinity of the quencher. The fluorescence intensity of
the reporter dye, as a result increases
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