Hydrolysis probes are DNA oligonucleotides that are labeled with fluorescent reporter molecule on the 5′ -end and a quencher molecule on the 3′-end

Hydrolysis probes are DNA oligonucleotides that are labeled with fluorescent reporter molecule on the 5′ -end and a quencher molecule on the 3′-end. Hydrolysis probes are designed complementary to the target and binds downstream of one of the primers (66) and increases specifictiy because of it binds specifically to a region of the DNA target (64). During amplification, the probe is degraded by the 5′- to 3′- exonuclease activity of the Taq polymerase. This cleavage of the probe separates the reporter molecule from the quencher, allowing the reporter molecule to emit fluroresence. When the reporter molecule and the quencher are close to each other, the fluorescence is suppressed (70)