Fig. 3.

ECS-CRISPR assay optimization: a) The ratio of crRNA to Cas enzyme, the concentration of Cas enzyme was 100 nM, b) Substrate RNA concentration. The concentration of Cas enzyme was 100 nM and the concentration of crRNA was 50 nM, c) Temperature dependency. The concentration of Cas enzyme was 100 nM, the concentration of crRNA was 50 nM, and the RNA substrate concentration was 2000 nM. The green value of the fluorescence image was analyzed by using the ImageJ software (NTC, non-template control reaction), d) Real-time fluorescence curve of RPA combined with fluorescent dyes, e) Influence of different RPA times on the real-time fluorescence curves of the detection. The concentration of Cas enzyme was 100 nM, the concentration of crRNA was 50 nM, the RNA substrate concentration was 2000 nM, and the incubation temperature was 39 °C. Bar graph data represented as the mean ± SD of three experimental replicates. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)