The Paleo- to Mesoproterozoic Birrindudu Basin is an underexplored frontier basin straddling the Northern Territory and Western Australia, and is a region of focus for the second phase of Geoscience Australia’s Exploring for the Future (EFTF) program (2020–2024). Hydrocarbon exploration in the Birrindudu Basin has been limited and a thorough assessment of the basin’s petroleum potential is lacking due to the absence of data in the region. To bridge this data gap, a comprehensive analytical program including organic petrology, programmed pyrolysis and oil fluid inclusion analysis was undertaken on cores from six drill holes to improve the understanding of the basin’s source rock potential and assess petroleum migration. Organic petrological analyses reveal that the primary maceral identified in the cores is alginite mainly originating from filamentous cyanobacteria, while bitumen is the most common unstructured secondary organic matter. New reflectance data based on alginite and bitumen reflectance indicate the sampled sections have reached a thermal maturity suitable for hydrocarbon generation. Oil inclusion analyses provide evidence for oil generation and migration, and hence elements of a petroleum system are present in the basin.