This study provides the first detailed lithostratigraphic and biostratigraphic constraints for improving stratigraphic resolution for hydrocarbon prospecting and exploration in the Tarim basin. A total of 49 stratigraphic units (38 formations and 11 members), ranging in age from the latest Devonian to Permian, are reviewed or redefined in terms of nomenclatures, lithology, age constraints, and lateral distributions based on the detailed field works or newly published data. Of these, the Piqiang Formation (new formation) is proposed to include the reefal carbonates of Asselian–Sakmarian age from the northern Tarim. The subsurface upper Paleozoic stratigraphic framework of the desert areas of the basin is also established for the first time. The high-resolution, basinwide stratigraphic correlations reveal that the sedimentation of the basin in the late Paleozoic was extremely uneven. Of these, the Famennian to Changhsingian successions are completely recorded in the southwestern margin areas of the basin. Here, five eustatic sedimentary cycles are well recognizable, suggesting the sedimentation was more eustatically controlled and little affected by local tectonism. The late Paleozoic successions of both Kalpin and Taklimakan regions are commonly interrupted by major hiatuses at various horizons, suggesting that the sedimentation was apparently modified by local tectonism. Of these, the northward movement of the Tarim block and its subsequent collision with the Yili microcontinent (part of the Kazakhstan plate) may be principally accountable for the discrepancy in the sedimentation of the various regions in the basin in the late Paleozoic.