Dual-Labeled Probes are the most common probe type for qPCR and are often referred to as hydrolysis probes.
A Dual-Labeled Probe is a single-stranded oligonucleotide labeled with two different dyes. A reporter dye is located at the 5’ end and a quencher molecule located at the 3’ end. The quencher molecule inhibits the natural fluorescence emission of the reporter by fluorescence resonance energy transfer (FRET). The illustration below depicts the mechanism.
The primer is elongated by the polymerase and the probe binds to the specific DNA template. Hydrolysis releases the reporter from the probe/target hybrid, causing an increase in fluorescence. The measured fluorescence signal is directly proportional to the amount of target DNA.
Our probes are provided in a format to simplify your experimental planning.
3Cyanine 3 alternative
5Cyanine 5 alternative