Remarkable observations on the adsorption and desorption mechanisms of single-stranded oligonucleotides and the hybridization of double-stranded DNA (ds-DNA) on a graphene oxide (GO) surface have been made using ensemble and single-molecule fluorescence methods. Probe and target DNAs labeled individually with fluorescence resonance energy transfer (FRET) pairs and having similar adsorption affinities toward the GO surface are used to provide detailed insights into the hybridization mechanism. Single-molecule FRET results reveal an "in situ" DNA hybridization mechanism, i.e., hybridization between the probe and target DNAs to form a ds-DNA, and simultaneous desorption from the GO surface thereafter. These results also demonstrate that the electrostatic interaction between DNA and GO is of little importance to the overall theory of interaction and the largest effects are from solvation forces, specifically the hydrophobic effect. This investigation improves the fundamental understanding of the DNA hybridization dynamics on the GO surface, opening new windows in the field of biophysics as well as in sensing and therapeutic applications.