The direct and selective assembly of deoxyribonucleic acid (DNA)-templated metal (e.g., Ag, Au, Cu, and Pd) nanowires (NWs) is a key technique for the application to electronic devices and nanowire-based biosensors. In this study, a new technique was developed to carefully control the interval of DNA-templated gold NWs (AuNWs) using surface-patterning techniques. The ??-DNA molecules were stretched and aligned along 3-aminopropyltriethoxysilane (APS) region formed uniformly by self-assembly and patterned by electron beam lithography process, and then treated by positively charged gold nanoparticles to form DNA-templated AuNWs. ??-DNA molecule was verified to be immobilized and stretched into parallel lines only on the surface of APS-coated parallel paths by surface-patterning technique, and it was possible to control the interval of DNA-templated AuNWs. Also, DNAs were confirmed to be assembled not on the bare SiO2 region but on the APS region defined by amine groups.