Blood genomic dna extraction kit

Genomic DNA was extracted from peripheral blood (Morning fasting whole blood) by using a Blood Genomic DNA Extraction Kit (Qiagen NV, Venlo, the Netherlands). And APOE genotype was determined by multiplex amplification refractory mutation system polymerase chain reaction (PCR). According to the methods previously described (23 (link)), these 169 subjects were divided into three groups, APOE E2 (ε2/ε2 and ε2/ε3, n = 25), APOE E3 (ε3/ε3, n = 111), and APOE E4 (ε2/ε4, ε3/ε4, and ε4/ε4, n = 33), and Supplementary Tables 1, 2 list the information about the gene distribution in detail. By using hexokinase method on an auto-analyzer (Dimension Xpand plus), we obtained the values of serum fasting blood glucose, triglyceride, cholesterol, high density lipoprotein, and low density lipoprotein.
The Research Ethical Committee of the affiliated mental health center of Shanghai jiaotong university school of medicine approved this study, and written informed consent was obtained from all participants before the study. All research processes was conducted in accordance with the principles of Declaration of Helsinki.

Genomic DNA was extracted from an EDTA-treated sample of the patients’ peripheral blood using the Blood Genomic DNA Extraction Kit (Qiagen, Germany). Apolipoprotein E (APOE) genotypes were determined by multiplex amplification refractory mutation system polymerase chain reaction (Jiang et al., 2018 (link)). The DNA sample of the proband was processed using WES. The details of the WES protocols and bioinformatic analysis have been described in previous publications (Li et al., 2017 (link); Jiang et al., 2019 (link)). In brief, an enriched DNA sample was sequenced on the Illumina Nova 6000 platform (Kangso Medical Inspection Co. Ltd., Beijing, China). All the variants were annotated using ANNOVAR software. The frequency of the variants in the general population was identified by the 1000 Genomes Project¹, the genome aggregation database², the ExAC database³, and the single-nucleotide polymorphism database⁴. SIFT⁵, CADD⁶, and PolyPhen-2⁷ were used to predict the possible functional changes caused by the variants. The potential variants were verified by Sanger sequencing. All the available family members, as well as the 500 healthy individuals, were sequenced to validate the confirmed variants.

Genomic DNA was isolated from peripheral blood using a commercial blood genomic DNA extraction kit (Qiagen). Sanger sequencing was performed to identify nonsynonymous variants, including frameshift, nonsense, deletion, or missense mutations in IRX3 (Gene ID: 79,191) exons with minor allele frequency (MAF) < 1%. Primers used to amplify the IRX3 exons are shown in Table S3.