WES Analysis
Genomic DNA was extracted from peripheral blood samples of proband and parents using standard procedures. The exonic regions and flanking splice junctions of the genome were captured using the Clinical Research Exome v.2 kit (Agilent Technologies, ). Sequencing was done on a NextSeq500 Illumina system with 150bp paired-end reads. Reads were aligned to human genome build GRCh37/UCSC hg19, and analyzed for sequence variants using a custom-developed analysis tool (Pezzani L Am J Med Genet A. 2018 Dec;176(12):2867-2871). Additional sequencing technology and variant interpretation protocol have been previously described (Pezzani L Am J Med Genet A. 2018 Dec;176(12):2867-2871). Coverage on target for the proband was ≥ 10x for 98% with a mean coverage of 203x.