In electrochemistry experiments on DNA-modified electrodes, features of the redox probe that determine efficient charge transport through DNA-modified surfaces have been explored using methylene blue (MBI) and Ru(NH3)(6)(3+) as DNA-binding redox probes. The electrochemistry of these molecules is studied as a function of ionic strength to determine the necessity of tight binding to DNA and the number of electrons involved in the redox reaction; on the DNA surface, MB+ displays 2e(-)/1H(+) electrochemistry (pH 7) and Ru(NH3)(6)(3+) displays 1e(-) electrochemistry. We examine also the effect of electrode surface passivation and the effect of the mode (intercalation or electrostatic) of MB+ and Ru(NH3)(6)(3+) binding to DNA to highlight the importance of intercalation for reduction by a DNA-mediated charge-transport pathway. Furthermore, in experiments in which MB+ is covalently linked to the DNA through a sigma-bonded tether and the ionic strength is varied, it is demonstrated that intercalative stacking rather than covalent sigma-bonding is essential for efficient reduction of MB+. The results presented here therefore establish that efficient charge transport to the DNA-binding moiety in DNA films requires intercalative stacking and is mediated by the DNA base pair array.