Graphene is an increasingly important nanomaterial exhibiting great promise in the area of nanotechnology. In this study, the azide-functionalized graphene derivative was synthesized as the 'click' reagent for preparation of polyvalent DNA-graphene conjugates, which provide an effective and stable platform to construct new functional nano-architectures. Assembled with Au nanoparticles, the prepared Au-DNA-graphene nanocomplex exhibits excellent stability that could prevent the nanocomplex from being destroyed by surfactants. Assembled with DNA tetrahedron-structured probes (TSPs), the nanocomplex displays outstanding sensitive electrochemiluminescence properties, which might be used as a biosensor for DNA detection. Therefore, this DNA-graphene conjugates could be explored as the assembly unit for advanced DNA nano-architectures in the field of DNA nanotechnology.