Exome Sequencing of DSB Formation Genes

Peripheral blood was collected from the patients, and the DNeasy Blood & Tissue Kit (Qiagen) was used to isolate the genomic DNA from leukocytes. Exome capture was carried out with SureSelect Target Enrichment System, and sequencing was performed on the Illumina platform (Illumina HiSeq). Reads were aligned against the National Center for Biotechnology Information (NCBI) hg19 reference human genome. Variants were called using ANNOVAR and Genome Analysis Toolkit. The variations in DSB formation genes, including SPO11, PRDM9, EWSR1, HELLS, MEI1, MEI4, IHO1, ANKRD31, REC114, and TOPOVIBL, were selected and classified according to American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) guidelines [14 (link)]. The pathogenic or likely pathogenic variations were confirmed by Sanger sequencing.