Massarray assay designer 3

In the fasting state, 3 mL blood sample was collected from each participant by a professional and stored with an anticoagulant tube of EDTA. Genomic DNA was then extracted from the participants’ blood samples by GoldMag-Mini Whole Blood Genomic DNA Purification Kit (GoldMag Co. Ltd., Xi′an, China) as the amplification template. The concentration and purity of genomic DNA were determined by Nanodrop 2000. Based on the 1000-genome project (https://www.internationalgenome.org/), we chose loci with minor allele frequencies (MAFs) >5% and Hardy-Weinberg equilibrium (HWE) >0.01 in the global population. Primers (amplification and extension) design were complete by Agena MassARRAY Assay Designer 3.1 (shown in the Supplementary Table 1). Based on Agena MassARRAY platform, genotyping of MMP17 SNPs were fulfilled. The sequencing data were sorted and analyzed by Agena Bioscience TYPER, version 4.0.

Based on the 1000-genome project (https://www.internationalgenome.org/) in the global population with minor allele frequencies (MAFs) >0.05 and Hardy-Weinberg equilibrium (HWE) >0.01, we selected variants of CYP19A1 gene. In addition, we applied online software-Agena MassARRAY Assay Designer 3.1 to design amplification primers and extension primers (Supplementary Table 1). By removing non-specific primers and ineffective primers, we finally selected four loci (rs28757157, rs3751592, rs3751591 and rs59429575) on CYP19A1 gene. Using our previously extracted genomic DNA as a template, we completed SNPs genotyping for CYP19A1 gene on the basis of the Agena MassARRAY nucleic acid mass spectrometry platform. Data sorting and analysis of the genotyping results were finished by Agena software Bioscience TYPER, version 4.0.