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. 2023 Nov 9;12(2):2276337. doi: 10.1080/22221751.2023.2276337

Figure 3.

Figure 3.

Optimization of the reaction conditions for the RT-RAA-CRISPR assay. A. Optimizing the reaction time of RT-RAA from 10, 20, 30, 40, 50 min. B. Optimizing the reaction temperature of RT-RAA from 25°C, 32°C, 35°C, 39°C, 42°C. C. Optimizing the reaction time of CRISPR–Cas13a from 20, 40, 80, 60, and 100 min. D. Optimizing the reaction temperature of CRISPR–Cas13a from 28°C, 33°C, 37°C, 41°C, and 45°C. E. Optimizing the concentration of Cas13a from 20, 45, 60, 100, and 120 nM. F. Optimizing the concentration of MgCl2 from 5, 10, 15, 20, 25 nM. G. Optimizing the concentration of crRNA from 100, 200, 300, 400 nM. H. Optimizing the concentration of T7 RNA polymerase from 0.2, 0.5, 0.8, 1 U/mL. The data are the mean ± s.d. for 3 technical replicates. ***: p < 0.001, ****: p < 0.0001.