Figure 2

Optimization and quantification strategies for the PIK3CA assays. (a) PIK3CA Assay n°1: ‘1D’ left panel showing optimized fluorescence signals produced by each probe in a simplex reaction; ‘2D’ center panel showing the polygon gating quantification strategy defined using a mixture of WT gDNA and MUT gBlocks for N345K, C420R, E542K, E545A/G/K/Q, Q546K/P/R and H1047L/R mutations; ‘3D’ right panel showing the cluster positions according to their relative FAM-HEX-Cy5 fluorescence signals. (b) PIK3CA Assay n°2: ‘1D’ left panel showing optimized fluorescence signals produced by each probe in a simplex reaction; ‘2D’ center panel showing the polygon gating quantification strategy defined using a mixture of WT gDNA and MUT gBlocks for E542K, E545K and H1047L/R mutations; ‘3D’ right panel showing the cluster positions according to their relative FAM-HEX-Cy5 fluorescence signals.