Recently, controlling assembly process in a precise manner has attracted increasing attentions in nanofabrication. Gold nanoparticles (AuNPs) modified with countable number of DNA strands, i.e., DNA discrete modified AuNPs, which bring AuNPs much more controllable manipulating possibilities have been playing an important role in this field. In this feature article, we will summarize recent progress on their preparation strategies and application in positioning assembly, which could benefit to the improvement of preparation methods of DNA discrete modified AuNPs and even other nanoparticles.