This paper is part of the larger body of genetic research being conducted on Nassau Grouper in the Caribbean. The authors identified ten polymorphic microsatellite loci for Nassau grouper (Epinephelus striatus) by cross-amplification of loci isolated in Gulf coney (Epinephelus acanthistius). Samples from three geographic localities were scored for these loci –Glovers Reef off Belize (n = 50), Little Cayman (n = 50) and Grammanik Bank in the U.S. Virgin Islands (n = 50). Screening samples yielded 8 to 27 alleles per locus with observed levels of heterozygosity ranging from 0.30 to 0.96. Markers will be used in a Caribbean-wide study of Nassau grouper to understand patterns of genetic connectivity, as well as to contribute to fisheries management and conservation.