We investigated population structure and genetic diversity for bobcats (Lynx rufus) in Michigan, USA, which are distributed throughout the upper peninsula (UP) and the northern half of the lower peninsula (LP) of Michigan. Specifically, we assessed the influence of natural and artificial barriers to dispersal on the genetic population structure of the bobcat across Michigan, as well as in each peninsula. We used 5 microsatellite markers and the statistical package STRUCTURE to identify populations and assign individuals to their population of origin. STRUCTURE identified one population in each peninsula, indicating that the UP and LP are genetically isolated by the Straits of Mackinac which divide the UP and LP. Despite a greater density of roads in the LP, we found no evidence that they have led to intra-peninsular population structure. Our results suggest that, from a genetic standpoint, management agencies do not need to be concerned about the fragmenting effects of roads when producing management plans for bobcats.