2.2 | WES
Genomic DNA was extracted from peripheral blood leukocytes using a
Wizard Genomic DNA Purification Kit following the manufacturer’s
instructions (Promega, Madison, WI, USA). SureSelect Human All Exon V5
(Agilent Technologies, Santa Clara, CA, USA) was used for library
preparation; and sequencing was performed on an Illumina HiSeq2500
platform (Illumina Inc., San Diego, CA, USA) to generate 2 × 100-bp
paired-end reads. Sequence reads were aligned against the Human
Reference Genome build GRCh37 using BWA 0.7.12; duplicated reads were
marked with Picard Tools 1.130; local alignment, base quality
recalibration and variant calling were conducted using the Genome
Analysis Tool Kit v3.4.0; and annotation and variant effect prediction
were performed with SnpEff v4.1g.