Dicer Fluorogenic Assay Kit

The Dicer Fluorogenic Assay Kit is a homogeneous 384-well assay designed to measure the double-stranded-RNA processing activity of Dicer for screening and profiling applications. The Dicer Fluorogenic Assay Kit contains enough purified Dicer, substrate, and assay buffer for 384 reactions.

Figure 1: Illustration of the Dicer Fluorogenic Assay method.
The assay relies on an RNA duplex imitating pre-miRNA, containing a conjugated fluorophore and quencher pair. Fluorescence of the dye is effectively quenched by the proximity of the quencher in the intact RNA duplex. The endoribonuclease Dicer processes the pre-miRNA into a mature 22 bp double-stranded miRNA by cleaving the 5’ end which is conjugated to the dye, releasing it from the miRNA. Separated from the quencher, the dye emits fluorescence with excitation/emission maxima of λ=647/ 665 nm. Dicer activity is proportional to the increase in fluorescence.