Nucleic acid bases are an important part of nucleic acids, and Raman spectroscopy is an important technique in studying on molecular structure. Therefore, it has a great significance for the research on the structural changes of nucleic acid macro-molecules and the interactions between nucleic acid molecules and small molecule that the Raman spectroscopy of nucleic acid bases are characterized. In this study, the molecular structures of adenine, guanine, cytosine, thymine and uracil were optimized based on the density functional theory method (DFT). The intra-molecular bond vibration frequency of these five nucleic acid bases were predicted by quantitative calculation, and the Raman spectroscopy of nucleic acid bases was characterized by corresponding the relation between the results of nucleic acid bases Raman spectroscopy in experiment and in theory. Moreover，the characteristic Raman peaks in the Raman spectroscopy of each nucleic acid bases were explanation in detail. It laid the theoretical foundation for the further structural information of nucleic acid molecules by the means of Raman spectra.