The specific polymorphic variant of the SLCO1B1 gene, the c.521 T > C SNP analyzed in this study (GenBank accession no. NC_000012.10), was performed by a polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) assay. PCR reaction volume was 25µL containing 1 µM of each assay-specific primers, 0.3 mM deoxynucleotide triphosphate (dNTPs) (Promega), 3 mM MgCl2 (Promega), 1.5 U Taq polymerase enzyme (Promega), 1 × PCR GoTaq Buffer Mix, water, and ≈ 1 µg genomic DNA. The PCR included 40 cycles at 94 °C for denaturation of the genomic DNA and activation of the Taq polymerase enzyme, 55 °C for annealing of the primers, and 72 °C for extension.
The PCR assay was performed using Tpersonal Thermocycler (Biometra), and finally, electrophoretic separation on 2% (W/V) agarose gel, with a running time of 90 min at 80 V in 1X TAE buffer (Tris–acetate-EDTA buffer; 40 mM Tris, 20 mM acetate, 1 mM EDTA), and visualization of the gel-separated PCR products with Green Safe (NZYTech) staining under UV light (AlphaImager, AlphaInnotech).
The PCR assay was performed using Tpersonal Thermocycler (Biometra), and finally, electrophoretic separation on 2% (W/V) agarose gel, with a running time of 90 min at 80 V in 1X TAE buffer (Tris–acetate-EDTA buffer; 40 mM Tris, 20 mM acetate, 1 mM EDTA), and visualization of the gel-separated PCR products with Green Safe (NZYTech) staining under UV light (AlphaImager, AlphaInnotech).