Fig. 2: Optimization of essential components of single-step RPA-CRISPR/Cas13a detection system. | Nature Communications

Fig. 2: Optimization of essential components of single-step RPA-CRISPR/Cas13a detection system.

From: Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device

Fig. 2

a Schematic of the mechanism of single-step RPA-CRISPR/Cas13a assay. b–g The end-point fluorescence intensity of the single-step RPA-CRISPR/Cas13a detection for the optimization of six decisive reaction conditions, including the rNTP concentration of 0.5, 1, 2, and 4 mM; the T7 RNA polymerase concentration of 0.5, 1, 2, and 4 U/µL; primer concentration of 50, 125, 250 and 500 nM; LwaCas13a concentration of 25, 50, 100 and 200 nM; crRNA concentration of 10, 25, 50 and 100 nM; the RNase H concentration of 0, 0.05, 0.1 and 0.2 U/µL. Mean ± s.d. for 3 technical replicates for (b–g).

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