FIGURE 4.

Overview of the iSCAN-OP detection assay. Schematic illustration of the iSCAN-OP detection assay. The diagram describes the addition of all necessary reagents: virus-specific RPA primers, reverse transcriptase, Cas12a protein, crRNA and HEX reporter to the tubes containing the RPA substrate, resuspended from the lyophilized pellet in resuspension buffer. After mixing, total RNA extracted from plant tissue and magnesium acetate is added to the reaction mixture. The reaction mixture is incubated at an isothermal temperature of 42°C for 20 min and the fluorescence signal is measured with the P51 Molecular Fluorescence Viewer.