By using the methods of hydrocarbon generation kinetics and carbon isotope kinetics, combined with geological background of natural gas pool formation, the generation and accumulation of natural gas from Yinan 2 gas pool was studied in Kuqa Depression of the Tarim Basin. Natural gas of Yinan 2 gas pool is mainly' derived from Middle and Lower Jurassic coal-bearing source rocks, and generally belongs to long time-accumulated gas. It is suggested that Yinan 2 gas is chiefly accumulated in the last 5Ma, its Ro ranges from 1.25% to 1.95%, and the loss rate of natural gas is about 25%-30%. This work not only complements and reduces the deficiency of formation model of natural gas pools which traditionally depends on the matching relation-ships between source rock, reservoir, cap rock, and trap, but also is a useful reference in the study of other gas pools.