Further validation of this approach can be inferred from the fact that we found similar frequencies of mutations in a number of genes previously detected by TCGA, including USH2A, FAT1, PLEC, SYNE1, PIK3CA, MLL2/KMT2A, MLL3/KMT2C, CASP8 COL6A6, and ZFHX4. There are a few other studies in the literature where ‘tumor-only’ protocols have been used. This evidence concerns the gene PIK3CA and neoplasm.