In this study, we reported development and validation of a 48-gene NGS panel that includes genes that are technically challenging for molecular profiling of myeloid neoplasms, such as CEBPA, CALR, and FLT3. The assay demonstrated good inter- and intra-assay precision for SNVs, indels including CALR 52 bp deletion, and FLT3-ITDs (Fig 2 and S5A and S5B Table). Here, CALR is linked to myeloid neoplasm.