The genotyping of CETP (rs5882, rs708272, rs3764261, rs1800775, rs2303790), RAGE (rs1800624 and rs1800625), and CYP4F2 (rs1558139) was carried out using the real‐time PCR. All single‐nucleotide polymorphisms (SNPs) were determined using TaqMan® Genotyping assays (Thermo Scientific).
The genotyping was performed using a Rotor–Gene Q real‐time PCR quantification system (Qiagen). Appropriate real‐time PCR mixtures of CETP (rs5882, rs708272, rs3764261, rs1800775, rs2303790), AGER (rs1800624 and rs1800625), and CYP4F2 (rs1558139) were prepared for determining SNPs.
A PCR reaction mixture (9 μl) was poured into each of 72 wells of the Rotor‐Disc, and then, 1 μl of matrix DNA of the samples (~10 ng) and 1 μl of a negative control (−K) were added.
The Allelic Discrimination program was used during the real‐time PCR. Then, the assay was continued following the manual provided by the manufacturer (www.qiagen.com , Allelic Discrimination). The program determined the individual genotypes according to the fluorescence intensity rate of different detectors: molecular marker labeled with VIC fluorescent dye was chosen for the X‐axis and a molecular marker labeled with FAM fluorescent dye was selected for the Y‐axis. These dye‐labeled probes were included in the TaqMan® Genotyping assays.
The genotyping was performed using a Rotor–Gene Q real‐time PCR quantification system (Qiagen). Appropriate real‐time PCR mixtures of CETP (rs5882, rs708272, rs3764261, rs1800775, rs2303790), AGER (rs1800624 and rs1800625), and CYP4F2 (rs1558139) were prepared for determining SNPs.
A PCR reaction mixture (9 μl) was poured into each of 72 wells of the Rotor‐Disc, and then, 1 μl of matrix DNA of the samples (~10 ng) and 1 μl of a negative control (−K) were added.
The Allelic Discrimination program was used during the real‐time PCR. Then, the assay was continued following the manual provided by the manufacturer (