The effect of various synthesis conditions in the fabrication of graphene using the exfoliation methods has been investigated. Graphite oxide and graphene fabricated by various synthesis conditions were identified by SEM and XRD. Graphite oxide was made from graphite by the chemical oxidation, and graphene was manufactured from graphite oxide by thermal exfoliation method. As a result, it is confirmed that graphite oxide was well formed from graphite, and the graphene could be obtained from graphite oxide. And it was found that the interlayer spacing between the graphene layers depended on the reaction time and particle size, regardless of the reaction temperature from 5℃ to 25℃.