Whole Exome Sequencing for Genomic Analysis

WES was performed on all the members of the four families using the Twist Human Core Exome Kit (Twist Bioscience, San Francisco, United States) according to the manufacturer’s protocol and sequenced with the Illumina NovaSeq 6000 platform. The BaseSpace pipeline (Illumina, San Diego, United States) and the TGex software (LifeMap Sciences, Alameda, United States) were used for the variant calling and annotation, respectively. Sequencing data were aligned to the GRCh37/hg19 human reference genome. Variants with a coverage lower than 10×, genotype quality (GQ) < 15, and gnomAD minor allele frequency (MAF) > 5% were excluded. WES results were interpreted according to ACMG guidelines 2015 (Richards et al., 2015 (link)).