Masterpure genomic dna purification kit

The procedure used for DNA isolation and genotyping was the same as in our previous study [13 (link)]. The MasterPure genomic DNA purification kit (Epicenter Technologies, Madison, WI, USA) was used for DNA isolation. SNPs of the PDGFA gene were genotyped using the TaqMan Predesigned SNP Genotyping Assay kits and the 7300 Real-Time PCR System (Thermo Fisher Scientific, Carlsbad, CA, USA). We re-genotyped 10–15% of samples to check the accuracy of genotyping. Repeatability of results reached 100%.
The polymorphisms for the analysis were selected from the Database of SNPs of the National Center for Biotechnology Information, U.S. National Library of Medicine [17 ]. We chose only SNPs where the frequency of the minor allele was ≥20%.
There were rs1800814 (G > A), rs2070958 (T > C), rs62433334 (C > G) variants. They are all located within the introns of the PDGFA gene.

Genomic DNA was extracted from peripheral leukocytes using a MasterPure Genomic DNA Purification Kit (Epicentre Technologies, Madison, WI, USA). The 11q23.3 locus polymorphisms, namely intergenic variants rs10750097 and rs1729410, and intron variant rs3741298 of the ZPR1 gene, were genotyped using a TaqMan^®Pre-designed SNP Genotyping Assay Kit (Applied Biosystems, Foster City, CA, USA). The 20 µL reaction mix consisted of 1 µL template DNA (15 ng/µL), 10 µL TaqMan^®Genotyping Master Mix (Cat. # 4371355), 1 µL probe (TaqMan^® Pre-designed SNP Genotyping Assay), and 8 µL deionized water. The probe was diluted (1:1) in a TE buffer (10 mM Tris-HCl, pH 8.0, 0.1 mM EDTA) before the reaction. PCR was performed according to the manufacturer’s specifications. Genotyping was successful in 90–98% of participants; the accuracy of this analysis was checked by re-genotyping 15% of the samples, and the repeatability of results was 100%. The LDlink Tool (https://ldlink.nci.nih.gov/?tab=ldmatrix, accessed on 27 July 2022) was used to create an interactive matrix of pairwise linkage disequilibrium statistics [11 (link)].

Genomic DNA was extracted from peripheral lymphocytes using the MasterPure genomic DNA purification kit (Epicentre Technologies, Madison, USA). The −930A>G polymorphism of the CYBA gene was genotyped using the TaqMan^® Pre-designed SNP Genotyping Assay (Applied Biosystems, Foster City, California, USA). The total volume of 20 μl of reaction mix included: 10 μl of TaqMan^® Genotyping Master Mix (Cat.# 4371355), 1 μl of probe (TaqMan^® Pre-designed SNP Genotyping Assay, Cat.# 4351376, C_11291925_10), 1 μl of DNA template (15 ng/μl) and 8 μl of deionized water. The probe was diluted with the TE buffer (1:1) before the reaction. The polymerase chain reaction amplification was performed according to the manufacturer’s specifications. Genotyping was performed using the 7300 real-time PCR system (Applied Biosystems).