Nimblegen seqcap ez human exome library v2

Exome sequencing was performed using a DNA sample from at least one affected individual from each of six families (Fig. 1a, c–f, h). In family ED238, exome sequencing was carried out in five family members, including three affected siblings and their unaffected parents. Sequence capture was performed in a solution using the Roche NimbleGen SeqCap EZ Human Exome Library v2.0 to target approximately 36.6 Mb of coding regions, and sequencing was carried out using the Illumina HiSeq or NextSeq 500 platforms. FASTQ files were aligned to human reference sequence (hg19) using the Burrows-Wheeler Aligner. The Genome Analysis Toolkit (GATK) was used for realignment of regions containing indels, recalibration of base qualities, and variant detection and calling,^{6 (link)} and variants were annotated using SeattleSeq137. Apparent novel homozygous variants were checked in public databases including dbSNP, 1000 Genomes, and Exome Variant Server (EVS), and in sequences of 400 normal individuals from the same ethnic groups. The FERMT1, FLG, and KRT14 genes were Sanger-sequenced as candidates in affected members of ED113, ED156, and ED236, respectively. Cosegregation of apparent pathogenic variants was tested by Sanger sequencing using DNA samples from the rest of the family members.