Abstract: The origin of dolostone in the Middle Jurassic Buqu Formation of the Plateau Basin has been a subject of prolonged debate. This study combines detailed petrological observations with analyses of Mg-C-O isotopes and elements to constrain the origin of dolostones in the Buqu Formation. Petrography and cathodoluminescence (CL) examination identified three types of matrix dolostones: very finely to finely crystalline dolostone (D1), finely to medium crystalline dolostone (D2), and medium to coarsely crystalline dolostone (D3). The analysis of the diagenesis sequence reveals that D1 originated from the dolomitization of grainstone in the early diagenetic phase, whereas D2 and D3 resulted from the recrystallization of D1 during the later burial phase. The presence of high Na (>100 ppm), low Fe (<1000 ppm), low Mn (<250 ppm), positive Ce anomaly, LREE enrichment, stable δ26Mg (−2.28‰ to −2.04‰), and δ13C (1.02‰ to 2.95‰) indicates that the early dolomitization fluid was oxidized seawater. As the crystal size increases (D1→D2→D3), the progressively rising Mn content and significantly negative δ18O (−10.72‰ to −7.81‰) suggest that the dolostone has experienced modification and alteration by buried pore water in the later stages. The fluctuations in relative sea level during the sedimentary deposition of the Buqu Formation were reconstructed through the utilization of Na, Sr/Cu, Sr/Ba, Rb/Sr, ∑REE, and δ13C. It was observed that the δ26Mg of dolostone closely mirrored the variations in sea level. The consistent trend of change confirms that sea level fluctuations control the formation and distribution of early dolostone. Frequent sea level rise and fall prompted the limestone deposited on the carbonate platform to be continuously transformed into dolostone, which accumulates over a long period to form large-scale thick dolostone. After the formation entered the burial stage, under the combined action of high Mg/Ca ratio pore water, high temperature, and high pressure, the early dolostone experienced the adjustment of burial dolomitization. This research offers a typical case study on the application of Mg-C-O isotope and elements to determine the origin of dolostone. This will aid in a more comprehensive understanding of the formation process of dolostone in ancient rock records.