Clinch Dace (Chrosomus sp. cf. saylori) is a newly recognized species of minnow with a restricted distribution in southwestern Virginia. Field sampling and genetic analysis support the hypothesis that Clinch Dace populations are small and fragmented. Analysis of neutral genetic markers shows that most Clinch Dace populations have undergone past bottleneck events and are being operated upon by random genetic drift. Bayesian cluster analysis showed that three out of the seven populations found in 2017 are distinct, while the other four show signs of more recent admixture. However, Fst values among streams were high and analysis of molecular variance indicated differentiation among populations in all streams. These findings support the view that these populations are genetically isolated. Effective populations sizes were low at most sites, enhancing the likelihood of loss of alleles to genetic drift. Low M-ratios, non-zero Fis values, and high degrees of relatedness among individuals indicate that some inbreeding is taking place. Habitat analysis did not identify variables affecting distribution or abundance of Clinch Dace populations. As the collection sites were targeted near known Clinch Dace occupied sites, it is likely that habitat variables known to impact Clinch Dace, such as conductivity, were within the species' range of tolerance. Results showed that Clinch Dace seem particularly resilient to sedimentation, corroborating earlier work showing a negative relationship between Clinch Dace abundance to sediment size. That is, small sediment size does not seem to have a negative impact on Clinch Dace abundance. Of all sites where Clinch Dace were found, only one culvert at one site was clearly perched and may present a barrier to upstream migration, a possibility which is supported by the genetic differentiation found among collections above and below that culvert. While this study demonstrates that selectively neutral genetic differentiation has taken place among Clinch Dace populations, it does address any local adaptation that may be taking place which would render translocations a risk for outbreeding depression. The findings of this study can inform conservation management in identifying possible sources of individuals for translocations among populations or for augmentation following captive breeding. / Master of Science / The Clinch Dace is a small, threatened minnow in the Clinch River basin that was unknown until 1999. Since then, research has addressed the biology, life history, and distribution of this fish. This study used data from selectively neutral genetic markers to analyze the population structure and degree of differentiation of Clinch Dace populations. My study sites were targeted at road crossings near known Clinch Dace populations to assess the effect of habitat fragmentation on Clinch Dace populations and to maximize the likelihood that I would collect enough genetic material for analysis. Genetic analyses showed that while there is some admixture among certain populations of Clinch Dace, there is differentiation at neutral genetic markers. This differentiation does not necessarily indicate adaptive variation among populations which could result in outbreeding depression should populations be mixed through translocations, but it is reason to proceed with caution. Road crossings were generally not found to be a cause of further population fragmentation in Clinch Dace, as demonstrated by genetic analysis and statistical analysis. Almost all of the occupied road crossing sites in this study were either embedded, free-flowing culverts that were not perched or small bridges, and these were not deemed to be obvious barriers to fish movement. The only exception was Hart Creek 2, where the culvert is slightly perched and Fst is high between populations in the upstream and downstream reaches. The results of this study will help to inform managers as to what conservation actions can be taken to improve population viability. One potential management action from this study could be the retrofitting of culverts that have become perched and are acting as barriers to Clinch Dace movement. Another potential conservation strategy is to translocate individuals from large population to small populations. The study determined: 1) which translocations might be acceptable based on the degree of genetic differentiation among populations, and 2) identified potential donor and receiving streams for translocations.