None of the CRC patients in our study carried both KRAS and BRAF mutations, supporting the hypothesis that KRAS and BRAF mutations occur in a mutually exclusive manner (Rajagopalan et al, 2002; Frattini et al, 2004; Ahlquist et al, 2008).
A similar correlation is observed for standardized heterozygosity using the number of segregating mutations (Supporting Figure S2).
This observation suggests that the concurrent CVs might be a mixture of SNPs and mutations, supporting the hypothesis that some CVS in dbSNP are truly phenotypic mutations, and some CVM in HGMD are disease-susceptibility SNPs.
Accumulation of roX1 ex40 RNA was unaffected by these mutations (Supporting Information, Figure S1A).
The known CMT genes accounted for the majority of our identified mutations supporting a correct clinical diagnosis.
Kinase-impaired BRAF mutants showed a significant association with concomitant activating KRAS or NRAS mutations, but not PIK3CA mutations, supporting the reported interaction of these mutations.
Sanger sequencing of a random sample of these mutations confirmed 35/35 mutations (Supporting Information, Table S1), indicating a very low false positive rate.
Since multiple nucleotide variants have been described at most of these sites, the test can detect 120 previously described mutations (Supporting Information Table S1).
Furthermore, a 5′-phosphorothiolate substitution, which creates a hyperactive leaving group, suppressed deleterious C75 mutations, supporting the role of C75 as a general acid.
In cases of family-specific mutations, supporting data from zebrafish functional (knock-down) experiments for genes such as MARS was obtained.
