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.