Surface-enhanced Raman scattering (SERS) substrates based on graphene and Ag nanoparticles hybrid structures with low cost, high uniformity were prepared by a standard process of immobilization of silver nanoparticles with 3-aminopropyltrimethoxysilane (APTMS). Thermal annealing was used for removing residual APTMS and adjusting the morphology of silver nanoparticles, and the effectiveness of this method was verified experimentally. The influence of annealing temperature, dipping duration, and APTMS volume on the distribution of Ag nanoparticles and Raman enhancement was investigated experimentally and analyzed in detail. Our samples were prepared under the preparation conditions of 10% v/v APTMS, dipping time of 48 h, annealing temperature of 450 degrees C, duration of 30 min, and Ar flow rate of 40 sccm. SERS activities with enhancement of 107 and relative standard deviation of <20% were observed using rhodamine 6G (R6G) as probe molecule with a concentration of 10(-6) M and 10(-7) M. (C) 2016 Elsevier B.V. All rights reserved.