Genetic Analysis
Next-generation sequencing was performed either by whole-exome sequencing using the SureSelect v.5 (no UTRs) exome reagent (Agilent Technologies) with variant filtering performed using the AgileExomeFilter program as previously reported42 or by a targeted panel-based resequencing of selected candidate genes using SureSelect RNA baits designed with the SureSelect Target Enrichment protocol (Agilent SureDesign wizard, Agilent Technologies) with filtering as previously reported.11 Sequencing was performed on a HiSeq 2500 in rapid run mode or HiSeq 2000 (Illumina). Sanger sequencing was performed to screen or confirm CCDC151 mutations in affected individuals and in other family members for segregation analysis, and details of the sequencing primers used are available on request.
Homozygosity mapping in family 71154 used either the whole-exome sequence data for affected individual 71154 II:2 or SNP genotyping data generated using the Affymetrix Genome-Wide Human SNP Array v.6.0 (Affymetrix) in both siblings 71154 II:1 and II:2. AgileMultiIdeogram, which is based on a previously published method43 modified to use software NGS-generated VCF rather than SAM files, was used to generate a visual output plus numerical chromosomal coordinates for autozygous regions of interest.