Figure 2

Pyrosequencing strategy and results for CYP2A6 *1 and CYP2A6*12 alleles. A) Schema of CYP2A6 *1 and CYP2A6*12 template sequences; arrows indicate forward PCR and sequencing primers, with degenerate nucleotide positions underlined. The two allele-specific nucleotides are boxed. B) Predicted histogram and pyrograms for CYP2A6*1/*1 (i), CYP2A6*1/*12 (ii), and CYP2A6*12/*12 (iii). Nucleotide dispensation order is shown at the bottom of pyrogram iii. Dispenses 2 and 3 interrogate the first allele-specific nucleotide (A in CYP2A6 *1 or T in CYP2A6*12), and dispenses 12 and 13 interrogate the second allele-specific nucleotide (C in CYP2A6 *1 or T in CYP2A6*12). Dispenses 1, 4, and 14 (circled) serve as negative background controls.