Understanding genetically encoded inherited differences in drug metabolism and drug targets offers the promise of minimizing adverse drug reactions and improving therapies. We have compared two real-time PCR-based methods, the TaqMan and LightCycler for the pharmacogenetic evaluation of CYP2B6 516G>T polymorphism. Both methods were found to be suitable for pharmacogenetic testing of CYP2B6 516G>T in the meaning of time consumption, accurate genotype calling, flexible reaction format, simple data analysis, and low cost per assay. The genotype success rate was similar, but the LightCycler procedure was less expensive in terms of cost per sample and shorter running time.