We isolated and characterised ten microsatellite loci from the reef manta ray, Manta alfredi. Loci were screened for 60 individuals from Japan. The number of alleles per locus ranged from 2 to 10 and observed heterozygosity ranged from 0.033 to 0.867. Cross-species amplification was successful in the sister species M. birostris (9/10 loci) and other mobulids, Mobula japanica (7/10), M. thurstoni (8/10) and M. tarapacana (5/10). These loci provide new tools for assessing population genetic structuring and genetic monitoring.