Genomic DNA was extracted from peripheral blood using a QIAsymphony DNA Mini Kit (Qiagen, Hilden, Germany). A custom capture panel (Dxome, Seoul, Republic of Korea) targeting coding exons and intron-exon boundaries of 497 genes related to hematologic disorders (Supplementary Table S1 ) was used. Prepared libraries were hybridized with capture probes and sequenced as paired-end reads (2 × 150 bp) using NextSeq 550Dx (Illumina, San Diego, CA, USA). NGS data analysis was performed with DxSeq Analyzer (Dxome). Single-nucleotide variants, small insertion and deletions, and copy number variants were identified [13 (link), 14 (link)]. All variants were classified and reported as a 5-tier system according to the American College of Medical Genetics and Genomics and the Association for Molecular Pathology guidelines [15 (link)]. Genes included in our panel that are associated with HHAs are classified according to their relevant phenotypes in Table 1 .
Genes included in our panel classified according to phenotype
| Phenotype | Genes included in the panel |
|---|---|
| Membranopathy | ANK1, SPTB, SPTA1, SLC4A1, EPB42, EPB41, PIEZO1, RHAG |
| Enzymopathy | G6PD, PKLR, AK1, AK2, ALAS2, GPI, NT5C3A, GCLC, GPX1, GSR, GSS, HK1, BPGM, TPI1, PFKL, PFKM, PGK1, ALDOA |
| Hemoglobinopathy | HBB, HBA1, HBA2, HBD |
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