2.3.2. Identification of CYP2D6 Alleles

Polymorphisms (\*1 and \*10) in the *CYP2D6* gene of individuals were determined using DNA sequencing analysis. The primers were designed and prepared using the primer3 software. The PCR was performed in 20 µL reaction mixture including 1 µL extracted DNA, 1 µL of 10 pmol of each primer (forward and reverse), and 17 µL autoclaved distilled water. The PCR program was composed of an initial denaturation at 94 ◦C for 3 min followed by 35 cycles of denaturation at 94 ◦C for 20 s, annealing for at 62 ◦C 30 s, and extension at 72 ◦C for 20 min. A final extension step was conducted at 72 ◦C for 5 min. The PCR products were then analyzed by direct sequencing using the ABI PRISIM BigDye Terminator Cycle Sequencing Kit and an ABI Prism 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).
