Ctenomys pearsoni (Pearson's tuco-tuco) is a subterranean rodent native to Uruguay. We tested the amplification pattern of 12 microsatellite loci, designed for C. sociabilis and C. haigi in a C. pearsoni population. DNA extractions were made from hair samples, and PCR amplification products were run on an ABI 3100 microcapillary gel. Eight loci were selected to form a highly polymorphic panel that could be used to efficiently screen populations of this species. In DNA from 35 tuco-tucos, the mean polymorphic information content value was 0.6536 and the mean expected heterozygosity was 0.7166. Paternity non-exclusion probabilities for seven independent loci were NE-1P = 0.0766 and NE-2P = 0.0108, and combined non-exclusion P(ID) was 6.2 x 10(-7). This panel of microsatellite loci has sufficient power to make inferences regarding group structure, mating strategies and evolutionary relationships among populations.