The spiny dogfish (Squalus acanthias) is the target of the largest U.S. shark fishery where stock size is monitored and assessed. Understanding and defining regional populations is essential to conservation efforts. Previous work on spiny dogfish in U.S. waters has concentrated on sampling from New England and the North Carolina/Virginia coast. The migratory habits and reproductive traits of these sharks have been described, but little can be found about the southernmost part of the population, including those occupying the coast of South Carolina and Georgia. This pilot study investigated the reproductive biology of the spiny dogfish in the southernmost region. Three family groups of spiny dogfish were sampled and five microsatellite loci were used to screen for multiple paternity. Both litter size and overall genetic diversity were consistent with values reported in other parts of the range. In addition, no evidence of multiple paternity or parthenogenesis was found in the three family groups. The analysis of additional samples will provide clarity about the southern population and contribute to effective population management.