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Jimusar sag, which lies in the Junggar Basin, is an important tight oil study area in China. However, the properties and origin of the crude oil and the geochemical characteristics of the tight oil from the Lucaogou Formation have not yet been studied. In the present study, 23 crude oil samples from the Lucaogou Formation were collected for analysis, including physical properties (density and viscosity), bulk composition, saturated hydrocarbon gas chromatography–mass spectrometry, and calculation of various biomarker parameters. Source rock evaluation and porosity permeability analysis were applied to the mudstones and siltstones. To analyze the hydrocarbon generation history of the Lucaogou source rocks, one-dimensional basin modeling was performed. The oil-filling history was also investigated by means of basin modeling and microthermometry. Biomarkers of suitable source rocks (total organic carbon content is >1%, summation of the free and volatile hydrocarbon and remaining hydrocarbon generative potential is >6 mg HC/g rock, and vitrinite reflectance is between 0.7% and 1%) were analyzed for oil–source correlation. The results indicate that low-maturity to mature crude oils originated from the source rocks containing terrigenous organic matter, which were deposited in a saline lake. The source rocks are of mainly type II kerogen. Oil in the lower section of the Lucaogou Formation is relatively dense and viscous and has elevated nonhydrocarbon content. In addition, a higher proportion of bacteria and algae is shown to have contributed to the formation of crude oil in the lower section when compared with the upper section of the Lucaogou Formation. Oil–source correlations demonstrate that not all mudstones within the Lucaogou Formation contribute to oil accumulation. Thin section observation indicates organic macerals surround the grains of some siltstones within the porous and permeable subsections, which proves the siltstones contribute to hydrocarbon generation. Crude oils from the upper and lower sections mainly originate from thin-bedded mudstones interbedded and siltstones within the porous and permeable subsections. The mudstones over or beneath the porous and permeable subsections consist of seals and prevent the vertical movement of oil by capillary forces. Despite being thicker, mudstones between the two porous and permeable subsections make no obvious contribution to the oil accumulation because of their low permeability and distance from the porous and permeable subsections, which is consistent with the principle of near-source charging for tight oil accumulations. Lucaogou oil has been charged twice, once from 235 to 210 Ma and again from 180 to 150 Ma, coinciding with oil generation time.