A nematode, Scottnema lindsayae, is the dominant metazoan found in soils of the McMurdo Dry Valleys, Antarctica. The distribution of S. lindsayae is patchy within and between these dry valleys; nevertheless, it is unclear to what extent these populations are genetically isolated. We investigated genetic diversity in this nematode using nuclear and mitochondrial gene sequences that encode ribosomal RNA. In 169 nematodes surveyed, only one variable site was found in each of two different expansion segments of nuclear rRNA. While most nematodes have only one sequence type, some nematodes were found to contain a mixture of both sequences. No fixed differences in nuclear sequences were observed between populations. This pattern of nuclear variation is most consistent with a single species of nematode defined morphologically as S. lindsayae. For mitochondrial DNA sequences, we found 10 variable positions defining 12 haplotypes among 188 nematodes surveyed. While all observed haplotypes are closely related, significant differences in haplotype frequencies were observed between geographically defined populations. The nuclear and mitochondrial variation suggests populations of S. lindsayae represent a single polymorphic species with some restriction of gene flow between geographic populations.