Graphene has been considered as a promising candidate to replace the traditional metallic oxide to prepare transparent conductive film based on its superior electrical conductivity and high visible light transmittance. However, so far no appropriate method is found to prepare high-quality and low-cost graphene in a large quantity. Therefore, the work aims to develop a technology to prepare the graphene based on fast electrochemical exfoliation of natural vein graphite and study the performance to exfoliate natural vein graphite and finally verify the feasibility to prepare the transparent conductive film by the graphene in volume production. The same electrochemical technology was used to exfoliate natural vein graphite and highly oriented pyrolytic graphiteand artificial graphite was used to prepare the graphene. The size and quality of the graphene exfoliated from natural vein graphite were investigated by OM, AFM, Raman and XPS. Then, the graphene exfoliated from natural vein graphite was compared with other two kinds of graphene and the graphene based on thermal/chemical reduction-oxidation reported in the literature. The transparent conductive film was prepared by the graphene exfoliated from natural vein graphite and its conductivity and light transmittance were measured. The graphene based on fast electrochemical exfoliation of natural vein graphite was mainly composed of 1—3 layers and the average horizontal size and thickness were respectively 5.9 μm and 2.4 nm. Through Raman and XPS analysis, the quality of such graphene could be compared favourably with that from highly oriented pyrolytic graphite by electrochemical exfoliation and better than that of artificial graphite and graphene based on thermal/chemical reduction-oxidation. Such graphene was used as raw materials to prepare the transparent graphene conductive film on the quartz by the interface self-assembly andthe subsequent transferring. At the light transmittance of 83.1%, the sheet resistance of the film decreased to 13kΩ/□, but improved greatly when compared with that of conductive film prepared by the graphene based on electrochemical exfoliation. The graphene prepared by electrochemical exfoliation with natural vein graphite as raw materials has large size, few defects and low degree of functionalization and can be used to prepare the transparent conductive film due to dense crystallization and high carbon contents of natural vein graphite.