High levels of fishing have resulted in declines among many of the oceans top predators, including the globally distributed tiger shark (Galeocerdo cuvier). Overexploitation of this species has led to declines in parts of its distribution, which may have important ecological consequences given the tiger shark’s trophic position as a large bodied, generalist predator. To assess the population genetics of this species, an enrichment protocol was used to isolate a suite of nine di- and tetra-nucleotide microsatellite loci within the tiger shark, which were subsequently characterized using genomic DNA derived from 40 individuals sampled from Hawaiian waters. The number of alleles per locus ranged from 3 to 23, and the average expected heterozygosities across loci ranged from 0.30 to 0.93. Analyses suggested a low frequency of null alleles across markers, and all loci conformed to both Hardy–Weinberg and linkage equilibrium at P > 0.01.