Currently the sex of Red-tailed hawks (Buteo jamaicensis) cannot be determined by in-hand methods. Males and females do not differ in plumage and overlap in size. During migration, we collected feather samples and morphological measurements from birds at four sites in the Western United States. Sex was determined for individual birds using sex-specific DNA markers and Polymerase Chain Reaction was used to identify these DNA markers. Through Discriminant Function Analysis, we created equations for determining the sex of Red-tailed hawks using in-hand measurements based on the DNA-determined sexes. We formed two equations, one for adults, which was 98% accurate, and one for hatch-year birds, which was 97% accurate. Our results will aid future studies looking at intra- and intersexual differences in the Western Red-tailed hawk.