We fabricated pattern of graphene oxide (GO) on a glass surface by chemical modification of the surface with the amino group and UV/ozone ashing using a microsieve as a photomask. Pattern of single-layered GO flakes were prepared with feasible processes at ambient pressure and temperature. The GO-patterned substrate was applied to the make arrays of supported lipid bilayer (SLB). SLB was formed at the GO-free region of the glass surface by the vesicle fusion method. Fluorescence recovery after photobleaching using flu-orescence microscopy showed that SLB in the array retained the membrane fluidity.