The invention discloses a conductive cotton fiber based on layer-by-layer self-assembled silver nanowire-graphene. The silver nanowire-graphene conductive cotton fiber is prepared by firstly, anionizing a cotton fabric, performing layer-by-layer self-assembling on silver nanowires and graphene oxide on the anionized cotton fabric, and finally reducing graphene oxide. Due to adoption of silver nanowire-graphene, the cotton fiber is excellent in conductivity, bacterium resistance, ultraviolet resistance, wrinkle resistance and scrubbing resistance, and has significant application values in fields of wearable equipment and flexible conductive devices.