DNA, the basic unit of heredity, is one of the most important biomolecules in the human body. In-depth structural insight of nucleotides is essential and has been actively pursued to provide new understanding of their biological function. With the current interest focusing on DNA and among the many developed methods and techniques to choose from, selection of the most appropriate experiments and methods has become increasingly difficult in order to obtain DNA solution structures with improved accuracy and reliability. Thus, careful decision must be made to maximize structural constraints with the greatest possible accuracy. This article reviews the application of nuclear magnetic resonance spectroscopy methods to the field of DNA structural determination that exists today with emphasis on conformation analysis.