We sourced archival CRC tumors (n = 8) that had been shown by the assessment of variant allele frequency (VAF) derived from NGS to contain possible subclonal driver mutations in KRAS, BRAF, or PIK3CA. Further samples (n = 3 ‘Ca-in-ad’ tumors) had been shown to carry subclonal KRAS mutations using the aforementioned pool of KRAS mutant probes. This evidence concerns the gene PIK3CA and colorectal carcinoma.