ERBB2 and cancer: Review of the VAF unique to each cell line and synthetic dilution mix demonstrated good agreement between the observed and expected AFs as established by the AmpliSeq Cancer Hotspot assay (ThermoFisher Scientific, Waltham, MA, USA, Pearson correlation of 0.9962 and 0.9975 for the validated SNVs/indel and HER2/neu copy-number, respectively) with an overall LoD of 10% (Figure 5b, Supplementary Table 9).