Diffusion Tensor Imaging (DTI) studies of non-human primates (NHP) are becoming increasingly common. Recently, many DTI analysis methodologies have been developed for human brain studies; however, few are directly applicable to NHP. A prerequisite for most statistical DTI analyses to localize WM differences across populations is to spatially normalize the individual scans to a representative template. Here, we report the development of a population-specific DTI template for young adolescent Rhesus Macaque (Macaca mulatta) monkeys using 271 high-quality scans. Using such a large number of animals in generating a template allows it to account for variability in the species. Our DTI template is based on the largest number of animals ever used in generating a computational brain template. It is anticipated that our DTI template will help facilitate voxel-based and tract specific WM analyses in non-human primate species, which in turn may increase our understanding of brain function, development, and evolution.