Massarray high throughput dna analysis

A DNA isolation kit, RelaxGene Blood DNA System (Tiangen, Beijing, China), was used for preparing genomic DNA following the recommendations of the manufacturer. Genotyping was performed by using MassARRAY high-throughput DNA analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (Sequenom, Inc., San Diego, CA, USA). The primers were designed by MassARRAY Assay Design software (Version 3.1). rs1333049 was genotyped using iPLEX Gold technology (Sequenom) followed by an automated data analysis with the TYPER RT software Version 4.0. Reproducibility of genotyping was confirmed by bidirectional sequencing in random 10% population, and the reproducibility was 99.1%.

Genotyping for TRPV1 SNPs rs222747 was performed in all patients. The MassARRAY Assay Design 3.1 software was used to design a single 20-multiplex reaction in which the SNPs of the TRPV1 gene was included. Genotyping was performed using iPLEX Gold technology (31 (link)) and MassARRAY high-throughput DNA analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (Sequenom).

Genomic DNA was extracted from 3 ml of venous blood samples using Blood DNA Maxi Kit (OMEGA, D3392) according to the manufacturer's protocol.
In the exploration cohort, genotyping was performed using MassARRAY high-throughput DNA analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (Sequenom, Inc., San Diego, CA, USA). 10% of samples were randomly selected for genotyping quality control. Subsequently, SNPs were genotyped using iPLEX Gold technology (Sequenom) followed by an automated data analysis with the TYPER® RT software version 4.0. In the enlarged validation cohort and HF, IS cohort, WNT3A rs752107 was genotyped by DNA sequencing after polymerase chain reaction (PCR).