Vacuette k3edta anticoagulant test tubes

At the time of study enrollment, venous blood samples (4 mL) for genotyping were taken into Vacuette K3EDTA anticoagulant test tubes (Greiner Bio-One, Kremsmünster, Austria) and stored at −80 °C. The manufacturer’s protocol for DNA isolation was followed (MagMAXTM DNA Multi-Sample Ultra 2.0 Kit, Thermo Fisher, Waltham, MA, USA), performed on a King Fisher Flex analyzer, and analysed by competitive allele-specific PCR (KASP-PCR) on the ABI 7500 analyzer (Applied Biosystems, Waltham, MA, USA USA); according to the finished reaction, a signal was read by the end-point method. Specific primers for the COMT polymorphism rs4689 were commercially prepared by LGC Biotechnologies (LGC Genomics, Teddington, UK). The KASP-PCR method was used for rs4689 COMT genotyping. According to the genotype, the wild-type homozygote (Val/Val) was defined as GG, the mutant homozygote (Met/Met) was defined as AA, and the mutant heterozygote (Val/Met) was defined as GA.

At the time of study enrollment, venous blood samples (4 mL) for genotyping were taken into Vacuette K3EDTA anticoagulant test tubes (Greiner Bio-One, Kremsmünster, Austria) and stored at −80 °C. The manufacturer’s protocol for DNA isolation was followed (MagMAXTM DNA Multi-Sample Ultra 2.0 Kit, Thermo Fisher, Waltham, MA, USA), performed on a King Fisher Flex analyzer, and analysed by competitive allele-specific PCR (KASP-PCR) on the ABI 7500 analyzer (Applied Biosystems, Waltham, MA, USA USA); according to the finished reaction, a signal was read by the end-point method. Specific primers for the COMT polymorphism rs4689 were commercially prepared by LGC Biotechnologies (LGC Genomics, Teddington, UK). The KASP-PCR method was used for rs4689 COMT genotyping. According to the genotype, the wild-type homozygote (Val/Val) was defined as GG, the mutant homozygote (Met/Met) was defined as AA, and the mutant heterozygote (Val/Met) was defined as GA.