Northern Yellow Bats (Lasiurus intermedius) exist in the southeastern United States and the southeastern counties of Texas along the coast of the Gulf of Mexico and extend into Mexico. The northern yellow bat is made up of two subspecies, L. i. floridanus and L. i. intermedius, which are differentiated by their size and the color of their pelage. There is a proposed line separating the two subspecies that goes through Atascosa County to Matagorda County. We used DNA sequence of the Cytochrome b gene to determine if there are two genetic lineages corresponding to the currently recognized subspecies. Samples were collected from individuals submitted to the Texas Department of State Health Services for rabies testing and supplemented with samples in the ASNHC. A total of 46 sequences originating in 12 counties and one sequence from an individual found in Mexico were included in a Maximum Likelihood analysis. Our results show that there were not two lineages recovered that correspond to the subspecies designations. We conclude that the specimens should be examined morphologically to confirm their subspecies designation and further molecular work should be conducted to test levels of gene flow across the range of this species.