Figure 4.

Action mode of dual hybridization probe. The dual hybridization probe consists of two hybridization probes, one brings a donor fluorophore at its 3′ end, and the other is labeled with an acceptor fluorophore at its 5′ terminus. After the denaturation phase, both probes attach to their target sequence in a head-to-tail arrangement during the annealing step. This causes the two dyes in close proximity to facilitate fluorescence resonance energy transfer (FRET). The donor dye in one of the probes transmits energy, facilitating the other one to dissipate fluorescence at a distinct wavelength (Adapted from Arya et al. [5]).