Neutron reflection has been used in combination with electrochemical and quartz microbalance techniques to characterize a mixed monolayer of thiolated hairpin ss-DNA and 4-mercaptobutan-1-ol monolayer self-assembled on gold before and after hybridization with a cDNA target. Neutron reflection has revealed the opening of the stem-loop configuration of the probe associated with helix formation. This change in conformation correlates to the modification of the electron-transfer resistance associated with the [Fe(CN)(6)](3-/4-) redox marker present in solution.