Pcr accessory set

A total of 1.5 µg of genomic DNA from each sample was bisulfite-treated with the an EZ DNA Methylation-Gold kit (Zymo Research, USA). The quality of the bisulfite conversion was controlled by using PCR products that had no methyl group. Bisulfite-treated DNA was amplified by a PCR Accessory Set (Sequenom, USA) followed by shrimp alkaline phosphatase (SAP) treatment, in vitro transcription (IVT) and RNase digestion by the MassCLEAVE Kit (Sequenom).

Genomic DNA was extracted from the stored EDTA anticoagulated-venous blood using a DNA purification kit (Puregene, Gentra Systems, Minneapolis, MN, United States). Then, the genotype of BDNF rs6265 was detected using the Sequenom method (Beijing Cnkingbio Biotechnology Corporation, Beijing, China). The following forward and reverse primers were used: 5′-ACGTTGGATGTTGT TTTCTTCATTGGGCCG-3′ and 5′-ACGTTGGATGGCTT GACATCATTGGCTGAC-3′, respectively. The target region was amplified using PCR Accessory Set (Sequenom, 11327). The PCR products were processed using iPLEX Gold reagents and purified using a SpectroCHIP Resin kit (Sequenom, 10117-2). MassARRAY^® Analyzer (Agena, 260000) was used to acquire data from the purified samples. No inconsistency was found when 10% of the samples were randomly selected and genotyped again.