“…A 25 μL reaction mixture contained the following reagents: 2.5 μL of 10x reaction buffer (100 mM tris-HCl, pH 8.0; 500 mM KCl; and 15 mM MgCl 2 ), 5 μL of 5 μM RNA reporter, 2.5 μL of 10 U/μL enzyme inhibitor, and 5 μL of 25 nM LwaCas13a nuclease. Then, 2.5 μL of 10 nM miRNA720 derivatives with different lengths (14,16,18,20,22, 24, 26, and 28 nt) and 5 μL of the 5 nM corresponding foldback crRNA sequence (42,40,38,36,34,32,30, and 28 nt, respectively) were added to the above reaction system. Finally, all reaction systems were incubated for 30−60 min at 25 °C in a real-time fluorescence quantitative PCR instrument, and fluorescence readings were measured every 30 s.…”