To estimate the stringency of the design and of these successive filters, we examined 71 kb of DNA using 518 amplicons that cover 4,446 mutated positions in the COSMIC database, with 5,271 known cancer mutations. The mutations in each sample were called using the algorithm trained on a calibration sample from the same run. Based on the amplicon design and filtering strategy presented above, we estimate that 70% of the targeted COSMIC positions can be assessed by our current method (Figure S10 in Additional file 1).
