Human Exome Sequencing Sample Clauses

Human Exome Sequencing. The study was approved by the Institutional Review Board (IRB) at Children's Mercy Hospital (CMH). Informed written assent/consent to participate in this study was obtained from all available family members. DNA was extracted from peripheral blood from the five living siblings and their parents. Exome sequencing was completed for V:14, VI:8, VI:9., and VI:11. DNA was not available from the four affected siblings who had died prior to moving to the United States. Library preparation was performed utilizing the KAPA Biosystems kit (KAPA Biosystems, Woburn, MA.) Enrichment using Illumina TruSeq Exome enrichment (Illumina, San Diego, CA) was performed according to manufacturers’ instructions. Samples were sequenced on an Illlumina HiSeq 2500 instrument with TruSeq v3 reagents, paired ~100 nucleotide reads. Alignment and variant calling was performed as previously reported(Xxxxx et al. 2014; Xxxxxx et al. 2015). Exome-enriched DNA was sequenced to a depth of 10.6Gb resulting in median target coverage of 67x. Variants were filtered to 1% minor allele frequency (MAF) in an internal database of 3974 samples, then prioritized by the American College of Medical Genetics (ACMG) categorization (Xxxxxxxx et al. 2015).