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Validation assays for G719S mutation detection in plasma samples. (A) Temperature gradient to determine the optimum annealing temperature; mutant positive events (top) and wild-type positive events (bottom) across the thermal gradient (57–67°C). (B) Two-fold dilution series of mutant DNA into wild-type DNA: concentration is shown in copies per microliter for mutant (blue) and wild-type (green) events and fractional abundance in percentage (orange). Error bars show 95% CI.
