In the case of APC, chromosomal deletions or frameshifting indels may be also responsible for APC inactivation and our methods may not adequately evaluate the genetic hierarchy of tumor suppressors such as APC. When we limit the APC mutations to those on a known mutation hotspot (APC:Q1387) accompanying TP53 mutations (four COADREAD cases), the CCF values of APC mutations were higher than those of TP53 mutations suggesting that APC mutation may have occurred earlier than TP53 mutation in those cases. Here, APC is linked to neoplasm.