Five CYP2A6 single nucleotide polymorphisms (SNPs) (rs1801272, rs28399442, rs28399433, rs1137115, rs28399435) were genotyped using the LGC Genomics Competitive Allele-Specific PCR (KASP) (lgcgenomics.com). The CYP2A6 copy number variant (CNV) was genotyped with TaqMan 5’ Nuclease Assays (Life Technologies). The CNV assay was run in duplicate, and genotype calls were made using CopyCaller software. The program PEDCHECK (O'Connell and Weeks, 1998) was used to examine Mendelian inheritance, and only individuals with no Mendelian inconsistencies were included in the analyzed sample. The metabolism metric was calculated based on the genotypes of the five CYP2A6 SNPs and the CNV using an algorithm described in Table S1 (adapted from Bloom et al., 2012).
