) by an enrichment by magnetic beads technique. One thousand three hundred and seventy-eight positive clones were obtained at the second screening by hybridization with Eight hundred and forty-six microsatellites were ultimately isolated from 705 positive clones. There were 632 (74.7%) perfect motifs, 114 (13.48%) imperfect motifs, and 100 (11.82%) compound motifs among the sequences. One hundred and thirty-seven primer pairs were designed according to the flanking sequences, and 100 pairs were synthesized. Thirty-five of the primer pairs (35%) showed polymorphisms in the wild population from the Hanjiang River. Twenty microsatellite markers were assessed for their genetic diversity, and a total of 212 different alleles were found. The average observed heterozygosity (e) was 0.797 5, and the polymorphism information content (PIC) was 0.777 7. The results suggested that the isolated microsatellites were valid for use in population genetic studies of grass carp