Abstract: A multi-chamber cartridge and a nucleic acid extraction module including the same are provided. The multi-chamber cartridge according to an aspect of the present invention may include a sample chamber including a first tube which is an elongated hollow type, a sample chamber body in which a mixing space is formed and one end of the first tube is disposed in the mixing space, a first pressure gasket which can be coupled to the inside of the first tube and is movable along the inner peripheral surface of the first tube, a first separation gasket which is disposed on one surface of the first pressure gasket, coupled to the inside of the first tube, and movable along the inner peripheral surface of the first tube, and a first plunger having one end coupled to the other surface of the first pressure gasket and pressing the first pressure gasket; and a cartridge body which includes an accommodating part in which the sample chamber is detachably accommodated.