The mechanism of interaction between lucidin (Luc) and DNA at physiological pH of 7.41) was investigated by ultraviolet-visible spectrum and fluorescence spectroscopy, using ethidium bromide (EB) as a probe. The effect of Luc on thermal denaturation and viscosity of DNA was also evaluated. In addition, the interaction between Luc and DNA was explored in a potassium iodide (KI)-mediated fluorescence quenching experiment. Luc was also subjected to electrochemical characterization on a glass carbon electrode, by cyclic voltammetry. Based on the curve obtained by cyclic voltammetry analysis, and the effect of EB on the interaction between Luc and DNA, the interaction was proposed to be mediated via intercalation. The binding ratio of Luc-DNA was determined to be 2:1, and the apparent molar absorption coefficient (ε) was determined to be 8.12 × 104 L/(mol?cm). The presence of Luc increased the thermal denaturation temperature and viscosity of DNA molecules, attenuated the fluorescence quenching effect of KI, and resulted in the display of similar electrochemical properties as EB. Under the conditions used in the present study, Luc was observed to interact with DNA molecules via intercalation.