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The pMini3bpUG, also known as Minihelix or Mh, can be used to measure the activity of RNase P as a minimal model substrate. The substrate utilizes a FRET mechanism in which an RNase P substrate is coupled to a 3 'fluorophore and a 5' non-fluorescent quencher. The cleavage and release of quencher molecules by RNase P enables detection of enzyme activity by measuring fluorescence emission over time, which is suitable for monitoring steady-state kinetics and high-throughput screening assaying.