An unknown occurrence of oil was detected near Avak Creek on Native lands on the North Slope of Alaska. Determining the source of oil was imperative for allowing stakeholders (Federal, State, and local government agencies and the landowner, an Alaska Native corporation) to make timely and informed decisions and mount a mitigation response, if required. The regional and local geological framework of the Avak Creek site was constructed using seismic surveys, well data, and basin modeling results, to identify local petroleum systems, map structural geometry and faults, define source rock thermal maturity distributions, and infer likely oil-migration pathways. Molecular hydrocarbon fingerprints (biomarkers, diamondoids, compound-specific isotopes) of the oil were compared to those of local and regional oil seeps, exploration well tests, and produced oils. Biomarker acid distributions characterized the history and extent of petroleum biodegradation. Integrating subsurface and geochemical parameters, the oil is interpreted to be a natural seep generated locally, predominantly from the Brookian Lower Cretaceous Hue Shale/gamma-ray zone, rather than an anthropogenic source of pollution. Results highlight sophisticated analytical technologies used to characterize complex, compositionally altered hydrocarbons. Results also advance our understanding of Brookian source rock distribution, subsurface petroleum migration pathways, and Arctic Alaska petroleum systems.