A genetic study was performed using NGS‐based whole‐exome sequencing (WES). WES was performed only in probands. Venous blood samples were collected from the probands and their families. DNA was isolated using DNeasy Blood and Tissue Kit (Qiagen) following the manufacturer's recommendations. In patient 1 and patient 2, WES was performed with SureSelectXT Human kit All Exon v7 (Agilent, Agilent Technologies), while in patient 3, Twist Human Core Exome (Twist Bioscience) was used. Then paired‐end sequenced (2x100bp) on HiSeq 1500 (Illumina). Bioinformatics analysis of raw WES data and variants prioritization were performed as previously described (Śmigiel et al., 2020 (link)). Prioritized NHLRC2 variants: (g.113876631G>T, NM_198514.4:c.442G>T, p.(D148Y) and g.113884318G>T, NM_198514.4: c.977G>T, p.(G326V) were validated in the probands and their families by deep‐amplicon sequencing (DAS) using Nextera XT Kit (Illumina) and sequenced on HiSeq 1500 (Illumina).
Runs of homozygosity (ROHs) were detected using the bcftools program as described previously (Narasimhan et al., 2016 (link); Smigiel et al., 2018 (link)). The reference group consisted of WES data from 559 unrelated Polish subjects from a local database.
Runs of homozygosity (ROHs) were detected using the bcftools program as described previously (Narasimhan et al., 2016 (link); Smigiel et al., 2018 (link)). The reference group consisted of WES data from 559 unrelated Polish subjects from a local database.
