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. 2015 May;17(3):284–292. doi: 10.1016/j.jmoldx.2014.12.003

Figure 5.

Figure 5

Sensitivity of the coamplification at lower denaturation temperature–droplet digital PCR (COLD-ddPCR) FAM/HEX ratio method for the detection of mutations in TP53 exon 8. A and B: FAM/HEX ratio values for serial mutant dilutions (100%, 10%, 5%, 1.25%, 0.63%, and 0%) of missense mutation p.R280K (located under the FAM-labeled probe) and mutant dilutions (50%, 5%, 1.25%, 0.63%, and 0%) for p.V274F (located under the HEX-labeled probe). C and D: FAM/HEX ratio values for serial mutant dilutions (100%, 10%, 5%, 1.25%, 0.63%, and 0%) of missense mutation p.R273H (located under the HEX-labeled probe) and serial dilutions (100%, 10%, 5%, 1.25%, 0.63%, and 0%) of p.R283C (located under the FAM-labeled probe). The lowest distinguishable mutation abundances in these experiments were 0.6% to 1.25% (P < 0.01). On the graph representing Taqman probes and DNA templates, the circles labeled on the left side of each probe are FAM (grey) or HEX (black). The circles labeled on the right side of each probe are quenchers (BHQ-1). The suns represent the release of FAM or HEX from the probe during COLD-ddPCR. The error bars represent variations among three independent detections.