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Real-Time Characterization of Ribozymes by Fluorecence Resonance Energy Transfer (FRET)
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1999
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Bioorganic ChemistryFret SubstratesBiochemistryMolecular Biological MethodNatural SciencesReal-time CharacterizationBioanalysisRibozyme ActivityExcitation Energy TransferMolecular BiophysicsRibozyme KineticsMedicinePhotophysical PropertySingle-molecule DetectionBiophysicsBiomolecular Engineering
More rapid than with conventional methods is the analysis of ribozyme kinetics upon use of FRET substrates. In these substrates a fluorophore and a fluorescence-quenching molecule lie in close spatial proximity. The intramolecular fluorescence quenching is neutralized upon cleavage, and the fluorescence intensity is a measure of the ribozyme activity. By automation and computer assistance the activity of ribozymes can be monitored under high-throughput conditions.