The size and the physical properties of graphene oxide sheets were controlled by changing the oxidation temperature of graphite. Graphite oxide (GO) samples were prepared at different oxidation temperatures of 20℃, 27℃ and 35℃ using a modified Hummers' method. The carbon-to-oxygen (C/O) ratio and the average size of the GO sheets varied according to the oxidation temperature: 1.26 and 12.4 μm at 20℃, 1.24 and 10.5 μm at 27℃, and 1.18 and 8.5 μm at 35℃. This indicates that the C/O ratio and the average size of the graphene oxide sheets respectively increase as the oxidation temperature decreases. Moreover, it was observed that the surface charge and optical properties of the graphene oxide sheets could be tuned by changing the temperature. This study demonstrates the tunability of the physical properties of graphene oxide sheets and shows that the properties depend on the functional groups generated during the oxidation process.