Quantstudio 3d digital pcr system life technologies amplifier

Blood samples for biochemical and pharmacogenetic studies were taken by antecubital venipuncture in vacutainer tubes with SiO₂ as a clot activator (to obtain serum) or with EDTA (to isolate genomic DNA by the standard phenol–chloroform method).
Genotyping of nine single-nucleotide polymorphisms (SNPs) of genes of the serotonergic system HTR1A (rs1423691, rs878567), HTR2A (rs6314), HTR3A (rs2276302, rs1150226), HTR2C (rs1414334, rs521018, rs498177), and SLC6A4 (rs16965628) was carried out using a MassARRAY^® Analyzer 4 mass spectrometer (Agena Bioscience ™) and a QuantStudio ™ 3D Digital PCR System Life Technologies amplifier (Applied Biosystems) using TaqMan Validated SNP Genotyping Assay kits (Applied Biosystems) based at The Core Facility “Medical Genomics”, Tomsk National Research Medical Center of the Russian Academy of Sciences.

Genotyping of 20 single-nucleotide polymorphisms (SNPs) in the ghrelin gene (GHRL: rs10490816, rs26312, rs26802, rs27647, rs35682, rs35683), leptin gene (LEP: rs2167270, rs3828942, rs10954173, rs4731426), leptin receptor gene (LEPR: rs1805094, rs1137101, rs1171276, rs1805134), and insulin-induced gene 2 (INSIG2: rs10490624, rs12623648, rs17047718, rs17587100, rs9308762, rs17047731) was carried out using a MassARRAY Analyzer 4 mass spectrometer (Agena Bioscience) and a QuantStudio 3D Digital PCR System Life Technologies amplifier (Applied Biosystems) using TaqMan Validated SNP Genotyping Assay kits (Applied Biosystems), based at The Core Facility “Medical Genomics” (Tomsk National Research Medical Center of the Russian Academy of Sciences). Selection criteria of the studied SNPs included a citation in the previous genetic studies of MetS components as described in the introduction, and a minor allele frequency (MAF) of > 5%. Basic information on these SNPs is described in Supplementary Table S2.

Blood samples were taken after 8 h overnight fasting in tubes containing EDTA and stored in several aliquots at −20 °C until DNA isolation using the standard phenol-chloroform method.
Inclusion criteria for SNP selection and genotype are described elsewhere [17 (link)]. Genotyping of eleven single-nucleotide polymorphisms (SNPs) in GRIN2A (rs7206256, rs1345423, rs8049651, rs9989388, rs7192557, rs9788936, rs9921541, rs11646587, rs1650420, rs11644461, rs4782039) and six SNPs in GRIN2B (rs220599, rs7313149, rs2192970, rs10845838, rs10772715, rs1805481) was carried out using a QuantStudio™ 3D Digital PCR System Life Technologies amplifier (Applied Biosystems, Waltham, MA, USA) using TaqMan Validated SNP Genotyping Assay kits (Applied Biosystems) based at The Core Facility “Medical Genomics”, Tomsk National Research Medical Center of the Russian Academy of Sciences.