Taqman snp predesigned genotyping assays

Genetic analyses were performed at the Institute for Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana as previously described [20 (link),22 (link)]. Briefly, genomic DNA was extracted from venous blood samples by using FlexiGene DNA kit 250 (Qiagen, Hilden, Germany). The LPA genotyping was performed by using Predesigned TaqMan SNP genotyping assays (C_30016089_10, C_25930271_10; Applied Biosystems, Waltham, MA, USA) and TaqMan™ Genotyping Master Mix (Applied Biosystems, Waltham, MA, USA) according to the manufacturer’s instructions. Analysis of LPA kringle repeats was performed by using custom TaqMan expression assay for exon 5 of the LPA gene. In brief, genomic DNA was diluted to 10 ng/µL with nuclease-free water. The PCR reaction mix contained 5 µL TaqMan Universal PCR Master Mix, 0.25 µL TaqMan Genotyping Assay Mix (40×), and 2.75 µL DNase-free water, to which we added 2 µL of the pre-diluted DNA. Genotyping PCR was performed with a QuantStudio 7 Flex Real-Time PCR System (Applied Biosystems, Waltham, MA, USA).

Genomic DNA was extracted from peripheral blood lymphocytes samples using Gentra Puregene Blood Kit (Qiagen, Italy) according to manufacturer's instructions.
Genotyping for the SNPs SOD2 A16V (rs4880), CAT −844 G>A (rs769214), and GPx1 rs1800668 C>T was carried out by real-time PCR allelic discrimination in a 7500 Fast Real-Time PCR instrument (Applied Biosystems, Foster City, California, USA), using Predesigned TaqMan SNP Genotyping Assays (Applied Biosystems; assay ID: C_1202883_20, C_850486_20, C_2548962_20). Reaction conditions and thermal profile were the same reported by Gugliandolo and coworkers [20 (link)].