Cancers arise as a result of genome instability and somatically acquired changes in the DNA of cancer cells. Some of these somatic mutations, known as driver mutations, induce growth and impaired differentiation leading to cancer development. Next-generation sequencing (NGS) enables us to identify hundreds of recurrent somatically altered genes. “Oncomine” focus assay (OFA) is commercially available NGS assay that enables the detection of variants in 52 cancer genes. OFA allows concurrent analysis of DNA and RNA to simultaneously detect hotspots, single nucleotide variants, indels, copy number variants (CNVs), and gene fusions in a single work flow using Formalin-fixed Paraffin-embedded (FFPE) samples. Six FFPE samples from five patients with various sarcomas (2 chondrosarcoma, 1 rhabdomyosarcoma, 1 fibrosarcoma and 1 Ewing sarcoma) were used in OFA. Four samples were needle biopsy specimens, and the other two samples were surgical excisional specimens. All patients were progressed after conventional chemotherapy or even though after high dose chemotherapy In total six cases, three were failed to analysis due to low cellularity, two cases showed no altered genes, and the other one case showed total seven CNV amplification (in MET, PIK3CA, FCFR4, FCFR1, GRAF, AR, and MYC). The case was a mesenchymal chondrosarcoma in left mandible. The disease progressed after three cycles of conventional chemotherapy. Then, operation was done and the excisional margin was positive to cancer. Now the patient is undergoing radiotherapy on primary site, and we plan to start targeted therapy for MET amplification. Molecular diagnostics have important implications on future treatment strategies. Furthermore, the identification of these molecular alterations driving the tumor enables clinical trials of novel agents. The OFA appears to be a suitable tool for fast, reliable, and cost-effective in molecular diagnostics. With as little as 10 ng RNA and DNA, OFA provides biomarker analysis focusing on the detection of genetic alterations. In this report, the number of cases is too small to assessing the effectiveness of OFA in sarcoma patients. Therefore, the result would be more convincing with a large number of cases and targeted mutations. Moreover, it is likely to have a customized panel for diagnosis and treatment among specific ages or types of cancer in near future.