What is the difference between static quenching and FRET?
Static (contact, ground state) quenching involves formation of a reporter-quencher dimer. The intramolecular dimer effectively decreases the concentration of the fluorescent reporter by creating a new, nonfluorescent reporter-quencher dimer with a unique absorption spectrum. FRET is a dynamic quenching mechanism that does not affect the probe’s absorption spectrum. Hybridization of the dual-labeled probe to its target or nuclease activity disrupts the reporter- quencher dimer, allowing the reporter to return to the state allowing fluorescence to occur.
