Limbless lizards of the genus Anniella are found in the western United States and Mexico. Until recently only two species were known, but four new species have since been described. Since these lizards are fossorial, not much is known about the nature of gene flow within species, or if gene flow occurs across species boundaries in regions of overlap. Since these lizards are of conservation interest, we isolated and developed nine tetranucleotide microsatellite loci for the recently described species Anniella alexanderae. We characterized the polymorphism of each locus in A. alexanderae, and then cross-amplified these loci in five other Anniella species. These nine loci have high observed levels of heterozygosity and polymorphism information content, and were in Hardy-Weinberg equilibrium within the A. alexanderae samples we tested, indicating that they will have high utility in assessing population genetic and demographic patterns within Anniella.