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

Figure 1.

Figure 1

A: Signal generation during enhanced ratio of signals for mutation scanning coamplification at lower denaturation temperature–droplet digital PCR (COLD-ddPCR). The system incorporates two hydrolysis/reporter probes that are complementary to the wild-type sequence. When the interrogated DNA region does not contain mismatches that may prevent the reporter probes to hybridize, both probes emit similar signals, translated into a 1:1 FAM/HEX ratio. B: If there is a mutation (G:C>A:T or G:C>T:A) present under one of the probes, the mismatch will affect the probe-target DNA binding, reducing the overall fluorescence of that particular probe, producing a >1 or <1 FAM/HEX ratio. 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.