Based on the comprehensive analyses of most data on stratigraphic temperature measurements, present geothermal gradient and heat flow of different units in the Tulufan Hami basin were detemined. The present geothermal gradient is 2.50℃/100 m and the heat flow value is 47.8mW/m2 in the basin. The geothermal gradient and heat flow is higher in the east and lower in the west respectively. The geothermal gradient and heat flow in the Tulufan Hami basin, which is higher than that of the Tarim basin and the Junggar basin, controlled by the following factors: crust thickness, basement topography and basin structure. Present temperature is an important control on oil gas generation of source rocks. The Taibei depression belongs to increasing temperature depression which is continuing burial, source rocks are on high stage of oil generation. In the Hami deprssiion and parts of Tuokexun depression the paleotemperature of source rocks is higher than that in the present. It was paleotemperature other than present temperature that controlled the oil generation.