_ Souring—or H2S generation—in the Bakken petroleum system (BPS) production is becoming problematic for oil producers and pipeline operators because of the sporadic occurrence and associated economic and HSE impacts. Paradoxically, crude oil produced from the tight Middle Bakken (MB) and underlying Three Forks (TF) reservoirs of the BPS has historically been light (40 °API to 50 °API) with low sulfur content (<0.2 wt.%), typically exhibiting nonexistent to low (<10 parts per million) H2S concentrations. Additionally, neither the MB nor the TF reservoirs have public records indicating the presence of H2S during drillstem tests (DST). However, based on previous work performed by the University of North Dakota Energy & Environmental Research Center (EERC), the frequency of H2S occurrence in BPS wells has increased from 2010 to 2023. Even though the observed H2S increases correlate with larger, multistage well stimulation and completion practices in the BPS, the unclear geographical and temporal trends make the exact mechanisms responsible for souring an open research question. At least five known mechanisms can potentially lead to H2S occurrence in the oil production stream of BPS wells, including: 1. Thermochemical sulfate reduction (TSR) 2. Bacterial sulfate reduction (BSR) 3. Source rock generation 4. Migration from deeper or shallower H2S reservoirs 5. Geochemical mechanisms related to reservoir stimulation work (Table 1). Understanding the mechanisms of H2S generation might reduce the risk of souring in new wells and enable more effective mitigation and management strategies in the wells already producing H2S. Samples Used in the Study This study is focused on oil and gas operations in the North Dakota portion of the Williston Basin and the H2S occurrences in the BPS, which includes two main oil-producing reservoirs: the MB and the TF. In the simplified stratigraphic sequence, the BPS comprises three distinctive lithofacies. From deepest to shallowest, those lithofacies are the Lower Bakken Shale (LBS), the MB reservoir, and the Upper Bakken Shale (UBS). Underlying the BPS is the TF, the upper part of which is the oil-rich TF reservoir, considered part of the BPS. The UBS and LBS are the source rocks for the oil in MB and TF reservoirs and consist of black, organic-rich, siliciclastic mudstones (i.e., shale). Directly overlying the BPS is the Madison Group, a carbonate-dominated Mississippian section that is subdivided into three formations—the Lodgepole (LP), Mission Canyon (MC), and Charles. The Birdbear Formation lies beneath the TF and on top of the Duperow Formation. Both the Duperow and Birdbear Formations produce oil and are dominated by carbonate deposits of shallow marine limestone and dolostones. A sampling program was developed to investigate areas of the BPS with varying levels of souring in the production stream and multiple organic-rich formations representing potential source rocks, including the UBS, LBS, LP, and MC. The samples were collected from locations and formations with different lithologies and maturity levels. Over 70 sour wells were identified for H2S sampling in western North Dakota.