Probe-based quantitative PCR (qPCR) uses real-time fluorescence from 5ʹ-3ʹ exonuclease cleavage of a fluorescently-labeled, target-specific probe to measure DNA amplification at each cycle of a PCR. Because probe-based qPCR is typically more specific than dye-based qPCR, it is often the foundational technology employed in qPCR diagnostic assays.
Probe designs vary but the most common type, hydrolysis (e.g., TaqMan®) probes, incorporate a 5’ reporter fluorophore and a 3’ quencher on a short oligonucleotide complementary to the target sequence. Fluorescence resonance energy transfer (FRET) prohibits emission of the fluorophore while the oligo probe is intact.
Probe-based qPCR allows multiple targets to be quantified in a single reaction (multiplexing) by using a unique fluorescent dye for each amplicon-specific probe.
SensiFAST™ Probe Kits have been developed by our partner Meridian Biosciences (Bioline) for fast qPCR and are designed for superior sensitivity and specificity with probe-detection technology, including TaqMan®, Molecular Beacon and Scorpions® probes.
A combination of the latest advances in buffer chemistry and PCR enhancers, in conjunction with an antibody-mediated polymerase promote rapid, consistently accurate detection of DNA and RNA targets from a broad range of sample types. To find the optimal SensiFAST kit for your real-time PCR instrument and application, you can also use this
Real-Time PCR Selection Chart.