AbstractThe positively charged nano‐ZnO and negatively charged natural DNA were alternately adsorbed on the surface of a gold electrode, forming (ZnO/dsDNA)nlayer‐by‐layer films. Valuable dynamic information for controlling the formation and growth of the films was obtained by cyclic voltammetry and electrochemical impedance spectroscopy. Differential pulse voltammetric (DPV) measurements showed that the electroactive probe methylene blue (MB) could be loaded in the (ZnO/dsDNA)nfilms from its solution, and then released from the films into Britton‐Robinson (B‐R) buffer. The complete reloading of MB in the films could be realized by immersing the films in MB solution again. However, after incubation in the solution of carcinogenic metal nickel, the damaged (ZnO/dsDNA)n films could not return to their original and fully‐loaded state, and showed smaller DPV peak currents. The results demonstrated that the DNA damage induced by the hydroxyl radical could be achieved by electrochemistry.