Figure 7 | Scientific Reports

Figure 7

From: Optimisation of robust singleplex and multiplex droplet digital PCR assays for high confidence mutation detection in circulating tumour DNA

Figure 7

Combination of FAM and HEX labelled probes to optimise mutant cloud separation in a PIK3CA multiplex assay. (a) Initial cloud distribution of the multiplex reaction was not optimal. (b) Addition of two H1047L mutant probes labelled with HEX and FAM dyes in 1:3 ratio moved the associated cloud to the right and in (c) 1:1 ratio, further moved the cloud to the right and lowered the channel 1 amplitude. (d) The final optimised cloud distribution in the presence of non-mutant DNA is shown. Neg – double negative droplets. Ex20 WT – PIK3CA Exon 20 WT Positive droplets. Ex9 WT – PIK3CA Exon 9 WT Positive droplets.

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