In this study, we isolated thirty-eight microsatellite loci from the draft genome of large yellow croaker, Pseudosciaena crocea. These loci were tested on 34 individuals from Zhangzhou prefecture city in Fujian. Most loci were highly polymorphic. A total of 214 alleles were detected with an average of 5.63 alleles per locus. The observed and expected heterozygosity ranged from 0.454 to 0.909 and from 0.503 to 0.918, with an average of 0.635 and 0.744, respectively. The PIC values of these loci ranged from 0.372 to 0.878, with an average of 0.712. Eight loci significantly deviated from Hardy-Weinberg equilibrium (HWE). Significant linkage disequilibrium (LD) was not found in these 38 loci. The newly identified polymorphic markers will contribute to the study of genetic diversity, population structure, and conservation of P. crocea.