Fig. 3: Development of a one-pot CRISPR/Cas12a-based assay.
From: Combination of MRI-based prediction and CRISPR/Cas12a-based detection for IDH genotyping in glioma
a Schematic of a one-pot method for IDH mutation status detection. Sequences of b IDH1-R132H and d IDH2-R172K crRNAs. Fluorescence heatmaps of different c IDH1-R132H and e IDH2-R172K crRNAs in Cas12a reactions. Time-course analysis of detecting 10 copies of f IDH1-R132H and g IDH2-R172K templates using our one-pot assay. Sensitivity of CRISPR/Cas12a-based detection of 1 × 104 templates with a gradient of h IDH1-R132H and i IDH2-R172K mutation rates. Data are presented as the mean ± SD (n = 3). ***p < 0.001, ****p < 0.0001 versus the WT group. j Sanger sequencing chromatograms of mutant dilutions. Minor components of the figure were obtained from BioRender.com.
