The invention discloses a preparation method of patterned graphene. The preparation method comprises the steps: a container with a patterned groove structure is used as a reaction container, a solution I and a solution II are respectively injected into the container, the liquid level is lower than the upper edge of a groove, the solution I forms a patterned thin film layer with nanoscale thickness on the surface of the solution II according to a pattern of the groove, and then the system is physically and chemically treated to make the solution I undergo a reaction, so as to obtain the patterned graphene, wherein the solution I is a graphene reaction raw material or a solution or suspension dissolved or dispersed with a graphene reaction raw material, and the solution II is a developing solution of the solution I. The method is simple and feasible, the cost is low, complex large instruments are not required, the reaction container is reusable and is easy to process, and the obtained patterned graphene has controllable pattern, has adjustable thickness, and is prone to industrialized preparation.