Whole Exome Sequencing Workflow

Whole exome sequencing was performed according to the referenced paper as follows [53 (link)]. WES was conducted on 500 ng of genomic DNA from the probands and their family members. Fragment libraries were prepared from the sheared samples by sonication and target enrichment was performed according to the manufacturer’s protocols (Agilent SureSelect QXT ALL Human Exon V6 Kit or Roche KAPA HyperExome Kit). Captured DNA was amplified followed by solid-phase bridge amplification and paired-end sequenced on Illumina NovaSeq 6000 (Illumina, Inc., San Diego, CA, USA). Alignment of reads to the human reference sequence (hg38 assembly) and variant detection were performed using DRAGEN 3.7.5 (Illumina, Inc.) with the alt-aware configuration. The variant annotation information was obtained from Variant Effect Predictor (version 100) and Jannovar (version 0.35) with dbNSFP 4.1a. The novel variants were filtered against 1000 Genomes (1000 genomes release phase 3, http://www.1000genomes.org/, accessed date: 28 July 2022), dbSNP (http://www.ncbi.nlm.nih.gov/projects/SNP/snp_summary.cgi accessed date: 28 July 2022), and the Genome Aggregation Database (gnomad.broadinstitute.org accessed date: 28 July 2022) [54 (link),55 (link),56 (link)].