Cutlassfish (Trichiurus lepturus Linnaeus 1758) is a commercially important fish in Korea. In recent years, the catch of cutlassfish in the coastal waters of Korea has significantly declined. Its genetic characterization has been little studied. To assist conservation and management efforts, we isolated and characterized 10 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. To characterize each locus, 30 individuals from a natural T. lepturus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except two, KTh9B and KTh22A, were polymorphic, with an average of 14.3 alleles per locus (range, 10 22). The mean observed and expected heterozygosities were 0.80 (range, 0.50 0.97) a 0.82 (range, 0.68 0.95), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at three loci (KTh6B, KTh10, and KTh16). This high variability indicates that these microsatellites may be useful for high-resolution studies of population genetics.