A simple and controllable method is developed to experimentally study the effects of defects on reduced graphene oxide (RGO) sheets for nanoelectronics application. First, a deterministic technique is developed to assemble a single layer graphene oxide sheet onto the gaps of microelectrodes by optimizing the dielectrophoretic parameters (10 V(pp) at 1MHz for 5s). This is followed by the utilization of atomic force microscopy-based mechanical cutting method to form line defects on RGO sheets. Based on these two procedures, the experimental studies of the effects of line defects on RGO are investigated, which provides an alternative approach to study the influence of defects on graphene. The electric transport measurement results show that the electrical performance of the defected RGO devices generally decrease due to Anderson localization, which supports the theoretical studies of the influence of defects on the electrical properties of RGO. (C) 2011 American Institute of Physics. [doi:10.1063/1.3665212]