- Course Introduction
- 01: Evidence & DNA
- 02: Forensic Biology
- 03: DNA Extraction & Quantitation
- Quantitation
President's DNA Initiative
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Advancing Justice Through DNA Technology
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Advancing Justice Through DNA Technology
DNA Analyst Training
Fluorescent Resonance Energy Transfer (FRET)
Home > DNA Extraction & Quantitation > Quantitation > Quantitative PCR > Fluorescent Resonance Energy Transfer (FRET)
Methods of fluorescent resonance energy transfer (FRET) technology involve a single probe where two dyes (the quencher and the reporter) are in close proximity. Due to this close proximity, an energy transfer occurs between the two dyes, suppressing the fluorescence of the reporter dye.09
Click here to read more about FRET at Wikipedia.com
Methods include:
- TaqMan® kit from Applied Biosystems uses a probe that hybridizes to the complementary target on the DNA strand. Polymerization cleaves the probe, releasing the reporter dye and resulting in fluorescence.10
Click here to view an animation about TaqMan kits.
- Molecular beacons are probes that form a hairpin loop with attached fluorescent reporter and quencher dyes. The fluorescent dye is suppressed while the probe is in this confirmation, due to the proximity of the dyes. Upon heating the probe (during the denaturation cycle of the PCR process) the hairpin structure is disassociated. This allows the probe (beacon) to bind to the PCR product in the subsequent annealing cycle. Binding of the beacon inactivates the quencher and releases fluorescence into the reaction.11
Click here to view an animation about molecular beacons.
