Fig. 4

Experimental results exploring the efficiency of primer depletion for Cas12a protein cleavage and validation of structure-specificity in the hairpin structure. a Fluorescence curves and endpoint bar charts comparing three parallel CRISPR reactions of RPA products with and without residual primers. Fluorescence was observed immediately under UV light after a 15-min reaction. Purified: RPA products with residual primers removed; Unpurified: RPA products without removing residual primers. b Fluorescence curves and endpoint fluorescence values of CRISPR reactions using crRNA with different numbers of mismatched bases compared to the original sequence when connected to a specific hairpin structure. The asterisk indicates significant differences as determined by two-tailed Student’s t-test (****P < 0.0001). ns denotes no statistical significance. NC/NTC negative control reaction