Fig. 3: Experimental observation of the working mechanism of DNACas.

a, b FAM terminal-labeled PO probe, PS probe, and PC&PS probe were incubated at room temperature (a) and 95 °C (b) with LbCas12a and subjected to polyacrylamide gel electrophoresis (PAGE) assay. DNA probes without incubation with Cas were also analyzed using electrophoresis as a control. c These analyses are identical to those shown in (a, b), except that the probe used is FAM mid-labeled PC&PS probe. d FAM terminal-labeled PO probe, PS probe, and PC&PS probe were incubated with LbCas12a and subjected to fluorescence polarization analysis. e Theses analyses are identical to those shown in (d), except that the probe used is a FAM mid-labeled PC&PS probe. The experiment results of fluorescence polarization were presented as mean ± standard error (n = 4 technical replicates). f PTS assay was employed to determine the melting curve of LbCas12a incubated with PO DNA1, PS DNA1, and PC&PS DNA1. In all experiments, UV represents 365 nm light irradiation for 1 min. P value was determined using an unpaired Student’s t-test. ***p \( < \)0.001, **p \( < \) 0.01, *p \( < \)0.05, ns-not significant (p\( > \)0.05). ‘M’ stands for marker. Source data are provided as a Source Data file.