Nanoscale step structures have attracted recent interest owing to their importance in both fundamental and applied research, for example in adsorption, in catalysis, and in directing nanowire growth. Here, we used a template-stripping method to obtain vicinal-like surface structures on grains of polycrystalline gold and investigated the effect of annealing temperature on the formation of these surfaces. Our results uncovered a correlation between the grain orientation angle (GOA) and the step periodicity and crystallographic direction on identical grains. The GOA was measured by determining the electron backscatter diffraction with respect to the sample's normal direction. Using scanning tunneling microscopy to examine identical grains, we found that their step periodicity decreases with increasing GOA.These results provide further understanding of the formation of periodically atomic gold steps at the gold/substrate interface, and thus might have promising potential for the directing growth of nanowires in microelectronics.