Extended Data Fig. 4: LoD analysis of RBD#3L, RBD#4L, and WT LwaCas13a in reactions supplemented with BSA and Triton X-100.
From: Engineered LwaCas13a with enhanced collateral activity for nucleic acid detection
a–c, Raw fluorescence data measured for 120 min, expressed as mean ± s.e.m. from four technical replicates. RBD#3L, RBD#4L and WT LwaCas13a were complexed with crRNA (cr1) and tested with 2.5, 1, 0.25, 0.1, 0.025 pM of the synthetic RNA (T1) in reaction buffer supplemented with 100 μg/mL BSA and 0.01% Triton X-100 (as in Supplementary Fig. 2b). The 0 pM target samples were used as non-target control (NTC). d, The bar plot represented the 120-min background-subtracted fluorescence expressed as mean ± s.e.m. from four technical replicates. Adjusted p values were calculated using two-way ANOVA with Dunnett’s test: **p = 0.0059, ***p = 0.0007, ****p < 0.0001, compared to background signal (non-target control) (see SourceData for adjusted p value). e, The velocities of each reaction within 60 min were obtained by linear regression from a–c and were plotted as functions of the target concentration. Outliers were identified by the ROUT method with a Q = 5%. Analytical LoD of three proteins was listed in Supplementary Table 2.
