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Ten highly variable microsatellite loci were performed in order to evaluate the genetic variability of wild and hatchery samples of half-smooth tongue sole Cynoglossus semilaevis. A group of 200 genotypes belonging to four wild samples, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ) and one hatchery sample, Mingbo (MB), were screened. All of the ten microsatellite loci screened in this study showed marked polymorphism. A total of 70 different alleles were observed over all loci. The number of alleles per locus ranged from 4.2 to 10.6.The average of expected and observed heterozygosity ranged from 0.67 to 0.82, and from 0.77 to 0.87, respectively. A total of 10, 16, 10, 11 and 5 unique alleles each were found in LZ, WH, QD,RZ and MB populations.The effective number of alleles varied from 2.87 for HSTS_7 to 6.83 for HSTS_h. The number of average genotypes ranged from 6.0 for HSTS_a to 15.4 for HSTS_h: AS compared with the wild populations, the hatchery population, MB, showed significant genetic changes such as fewer alleles per locus (P＜0.05), a smaller number of low frequency alleles (P＜0.05), a small number of unique alleles and a small number of genotypes (P＜0.05), all indicative of a reduction in genetic diversity.