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Acanthogobius ommaturus,a fish species of the Family Gobiidae,is a marine commercial fish perched on the bottom of seawater.In this study,Illumina high-throughput sequencing technology was applied to obtain the candidate microsatellite markers ofA.ommaturus.A total of 4 746 microsatellite-rich fragments were found,of which 4 542 microsatellites are with primer fragments,containing 971 dinucleotide sequences,2 643 trinucleotide sequences,569 tetranucleotide sequences,406 pentanucleotide sequences,and 212 hexanucleotide sequences.Based on the results of high-throughput sequencing,a total of 141 pairs of the microsatellite primers were designed and screened.And then 24 polymorphic primers were finally obtained by polyacrylamide gel electrophoresis.In total,271 alleles were detected in the 24 pairs of primers.The number of alleles for different primers ranged from 5 to 19.The average number of effective alleles (Na) was 11.292;the average observed heterozygosity (Ho) of the 24 pairs of primers was 0.665,the average expected heterozygosity (He) was 0.880,and the average polymorphic information content was 0.846.All sites were highly polymorphic (PIC＞0.50).