The status of desert bighorn sheep (Ovis canadensis nelsoni) populations in the mountains around Death Valley was first evaluated in 1938, shortly after designation of Death Valley National Monument. However, the most comprehensive evaluation of bighorn sheep in the region was conducted by Ralph and Florence Welles during 1955-1961. They documented patterns of use at water sources and other focal areas around Death Valley and roughly estimated numbers of bighorn sheep from observational data. Data collection on bighorn sheep in the area since that time has lacked a regional approach needed to address metapopulation questions.From 2011-2013, we evaluated bighorn activity at important water sources and other likely locations around Death Valley using remote cameras and observations of tracks, beds, sign, and bighorn sheep, and non-invasively collected genetic samples (fecal pellets and bones). Where possible, we revisited many of the water sources and other locations originally investigated by Welles and Welles (1961) and earlier researchers. We extracted DNA from fecal pellets, carcass tissue samples, and blood samples archived from earlier captures and genotyped them using highly variable genetic markers (15 microsatellite loci) with sufficient power to distinguish individuals and characterize gene flow and genetic structure. We also analyzed DNA samples collected from other bighorn sheep populations extending north to the White Mountains, west to the Inyo Mountains, south to the Avawatz Mountains, and southeast to the Clark Mountain Range, Kingston Range, and Spring Mountains of Nevada. We estimated genetic structure and recent gene flow among nearly all known populations of bighorn sheep in and around Death Valley National Park (DEVA), and used assignment tests to evaluate individual and population-level genetic structure to infer connectivity across the region. We found that bighorn sheep are still widely distributed in mountain ranges throughout DEVA, including many of the areas described by Welles and Welles (1961), although some use patterns appear to have changed and other areas still require resurvey. Gene flow was relatively high through some sections of fairly continuous habitat, such as the Grapevine and Funeral Mountains along the eastern side of Death Valley, but other populations were more isolated. Genetic diversity was relatively high throughout the park. Although southern Death Valley populations were genetically distinct from populations to the southeast, population assignment tests and recent gene flow estimates suggested that individuals occasionally migrate between those regions, indicating the potential for the recent outbreak of respiratory disease in the southern Mojave Desert to spread into the Death Valley system. We recommend careful monitoring of bighorn sheep using remote cameras to check for signs of respiratory disease in southeastern DEVA and ground surveys in the still-understudied southwestern part of DEVA.