Whole-exome sequencing

Genomic DNA was extracted from whole blood and sheared with an S2 Focused-ultrasonicator (Covaris). An adaptor-ligated library was prepared with the TruSeq DNA Sample Prep Kit (Illumina). Exome capture was performed with the SureSelect Human All Exon V5 Kit (Agilent Technologies). Paired-end sequencing was performed on the HiSeq 2500 System (Illumina) generating 100-base reads. The sequences were aligned with the GRCh37 build of the human genome reference sequence, with the Burrows-Wheeler Aligner (78). Downstream processing and variant calling were performed with the Genome Analysis Toolkit (79), SAMtools (80), and Picard tools (http://broadinstitute.github.io/picard/). All variants were annotated with in-house annotation software.