Whole-exome sequencing of blood
Genomic DNA was extracted using the QIAamp DNA Blood Extraction Kit (Qiagen, Germany) and fragmented to 150 -200 bp with the Covaris S2 ultrasonicator (Covaris, USA). Libraries were prepared with the NEB Next DNA Library Preparation Kit-Illumina (NEB, USA) and quality controlled with the Nanodrop 2000 and Agilent 2100 Bioanalyzer (Agilent Technologies, USA). WES was performed using the WES kit (MyGenostics, Beijing, China), which was applied to capture exon regions and their upstream and downstream 50 bp. The captured regions were sequenced with 150 bp read length using the Illumina HiSeq X ten platform. Low-quality data and junctions were removed from the sequencing data. The BWA software was used to align to the reference genome (hg19 version) and statistically analyze data such as sequencing depth, homogeneity, and probe specificity. The GATK software was used to compare sample data for polymorphic sites, and SNPs and InDels were statistically analyzed. The ANNOVAR software was used to annotate the data with all SNPs and INDELs, such as 1000 genome, ESP6500, Exome Aggregation Consortium (EXAC) and EXAC-EAS (EXAC ~4000 East Asian data)