Species reintroductions are successful when established populations maintain both demographic stability and genetic diversity. Such a result may be obtained by ensuring both structural habitat connectivity and genetic connectivity among reintroduced and remnant populations. Nevertheless, prezygotic barriers such as assortative mating can prevent the flow of genetic material between populations, even when migration between populations is high. Limited gene flow may be particularly relevant for reintroductions that were sourced either from captive-bred populations or from disparate locations in the wild. American martens (Martes americana) have been reintroduced repeatedly in the Upper Midwestern United States in an effort to establish self-sustaining populations. We quantified levels of genetic diversity within and spatial genetic variance among four marten populations during two time periods separated by 10 years. Spatially informed and naïve discriminant analysis of principal components were used to assign individuals to populations. Results indicate that heterozygosity declined and inbreeding coefficients increased between the two collection periods, while genetic structure among populations also increased. Data are consistent with assortative mating contributing to reapportioning of genetic variation. Population assignment tests show that migration among populations is apparent, but admixture (based on cluster membership probabilities) is low and declined over time. Specifically, martens may be successfully dispersing between populations but a lack of admixture indicates a lack of reproductive contributions to genetic diversity by migrants. Because marten reintroductions in this region are well-documented and well-monitored, lessons can be derived from results to inform future reintroductions. We encourage a careful balance of supplementing genetic diversity via augmentation while avoiding translocation of animals from disparate populations that may result in reproductive isolation of migrants. In combination with the maintenance of a functionally connected landscape, this strategy would maximize the likelihood of a successful reintroduction in terms both of demography and genetics.