In the northern Perth Basin (Western Australia), the Early Triassic Kockatea Shale is the primary petroleum source rock. Possible source rocks in the Northern Carnarvon Basin are more varied and include the Upper Jurassic Dingo Claystone as well as the Early Triassic Locker Shale. Biomarker analyses were conducted on petroleum samples from these basins to understand the nature of the petroleum systems. Many of the analysed petroleum samples contain carotenoids (okenane, chlorobactane and isorenieratane) derived from photosynthetic sulfur bacteria, suggesting that their source rocks were deposited under conditions of photic zone euxinia (PZE) and/or derived from microbialites. In the northern Perth Basin, the major lithofacies contributing to the source rock are dark coloured mudstones deposited under PZE conditions and/or derived from microbialites. In the southern Perth Basin, the potential source rock is either Permian, Jurassic or Cretaceous in age as indicated by the low concentrations or absence of carotenoids and the Triassic biomarker n -C 33 alkylcyclohexane. There is also a possibility that the Lower Triassic Locker Shale is the source rock of petroleum in the Tubridgi field on the Peedamullah Shelf of the Northern Carnarvon Basin, based on the similarity of biomarkers to Perth Basin petroleum sourced from the Kockatea Shale. However, the possibility of charge from the Upper Jurassic Dingo Claystone cannot be entirely excluded. Supplementary material: biomarker dataset is available at https://doi.org/10.6084/m9.figshare.c.6452153