We isolated and characterized a total of 33 microsatellite loci from the Patagonian sprat Sprattus fuegensis. Loci were screened in 24 individuals from the inshore waters of the Aysén Fjord, Chile. The number of alleles per locus ranged from 7 to 24, observed heterozygosity ranged from 0.217 to 0.875, and the probability of identity values ranged from 0.006 to 0.133. These new loci will provide tools for examining population genetic structure, estimating effective population size and provide information to fisheries management.