We surveyed population-level sequence variation in part of the mitochondrial control region for three species including eight subspecies of Cepphus guillemots (Charadriiformes: Alcidae) to test specific predictions about mechanisms of population differentiation. We found that sequences of spectacled guillemots (C. carbo) were more closely related to those of pigeon guillemots (C. columba; both found in the Pacific Ocean) than to those of black guillemots (C. grylle; Arctic and Atlantic Oceans), despite dissimilarities in plumage between spectacled guillemots and the other species. Distributions of species and timing of divergence events suggest that speciation involved allopatric and microallopatric populations isolated by Pleistocene glaciers. Control region sequences were significantly differentiated among populations within species and suggest that gene flow is low; however, populations are probably not in genetic equilibrium, so these results probably reflect historical isolation of colonies. In contrast, phylogenetic relationships among sequences within species were poorly resolved, probably because of a combination of incomplete lineage sorting and contemporary gene flow. Indices of genetic diversity provided no suggestion of recent bottlenecks in most populations, although two populations apparently underwent recent severe bottlenecks. Genetic divergence among populations was not correlated with geographic distance, which argues against isolation by distance. Results of these analyses, combined with breeding distributions and timing of divergence events, suggest that populations diverged during isolation in glacial refugia. Our results are consistent with earlier hypotheses posed by Storer and Udvardy.