The invention relates to self-assembly graphene nanometer thin film and a preparing method and application thereof. The method specifically comprises the following steps of 1, providing first dispersing liquid, a second solvent and a substrate, wherein the first dispersing liquid comprises a first solvent and a graphene material dispersed in the first solvent; 2, dropwise adding the first dispersing liquid into the second solvent at a first speed, and based on the Marangoni effect, obtaining a thin layer of the graphene material on the liquid level of the obtained mixed liquid; 3, transferringthe obtained thin layer of the graphene material in step 2 into the substrate, and drying the obtained product to obtain the self-assembly graphene nanometer thin film. The invention further discloses the self-assembly graphene nanometer thin film prepared by means of the method and the application of the self-assembly graphene nanometer thin film. The method is quick, efficient, low in cost, safe, environmentally friendly, free of limitation on the substrate and capable of preparing the graphene nanometer thin film on a large scale. The graphene nanometer thin film is good in uniformity, high in light transmittance and excellent in electric conductivity.