The invention relates to a graphene-organic material layered assembly film and a preparation method thereof. The preparation method comprises: using a graphene material and an organic material as raw materials, utilizing interaction of static electricity, hydrogen bonds, coordinate bonds or charge transfer and the like between the graphene and the organic material, and superposing films layer by layer through the film preparation methods such as spin coating, spraying, dipping, lifting and pulling and the like to prepare the film, wherein the thickness of each layer of the film can be controlled between 10 nanometers and 2 millimeters according to requirement. The layered assembly film and the preparation method have the characteristics that multilayer film materials with different functions are prepared by utilizing unique electric, magnetic, mechanical and chemical properties of the grapheme, and can be used as biomaterials, conductive materials, electromagnetic shielding and wave absorbing materials, photovoltaic materials, electrode materials, film filtering and separating materials, and the like to be applied to chemistry and chemical industry, biology and precision instruments, and manufacture of micro electrons, machinery and aviation and aerospace devices according to the selected different organic materials.