Different from most tight oil reservoirs, the tuffaceous tight oil of the Tiaohu Formation is not in situ oil, and no close contact exists between the source rock and reservoir in the Malang sag (Santanghu Basin, China). This study determined the mechanism of hydrocarbon accumulation of this tuffaceous tight oil reservoir through an integrated analysis of oil–source rock correlation, reservoir characteristics, and rock wettability combined with a comprehensive analysis of geological conditions. An oil–source rock correlation using biomarkers and stable carbon isotopes shows that the crude oil originated from underlying source rocks in the Lucaogou Formation. The oil in the tuffaceous tight reservoir is not indigenous but has migrated over a long distance to accumulate in these reservoirs. Faults and fractures that developed at the end of the Cretaceous are the oil migration pathways. Vitric and crystal-vitric tuffs constitute the main rock types of the tuffaceous tight reservoir. Matrix-related pores in the tuffs mainly comprise interparticle pores between minerals and dissolution intraparticle pores formed by devitrification. The adsorption of polar components of the oil generated from original organic matter in the tuff leads to wettability of lipophilicity, which is the main reason for hydrocarbon charging and accumulation. To our knowledge, this is the first comprehensive study reporting this finding.