The binuclear structure of Fe2(DTPB)(μ-O)(μ-Ac)CI(BF4)2 (DTPB = 1,1,4,7,7-penta (2′-benzimidazol-2-ylmethyl)-triazaheptane, Ac = acetate) was characterized by UV-visible absorption and infrared spectra and NMR and ESR. The binding interaction of DNA with the diiron complex was examined spectroscopically. Supercoiled and linear DNA hydrolytic cleavage by the diiron complex is supported by the evidence from anaerobic reactions, free radical quenching, high performance liquid chromatography experiments, and enzymatic manipulation such as T4 lig...