DNA-directed assembly of nano-objects as a means to manufacture advanced nanomaterial architectures has been the subject of many studies. However, most applications have dealt with noble metals as there are fundamental difficulties to work with other materials. In this work, we propose a generic and systematic approach for functionalizing and characterizing oxide surfaces with single-stranded DNA oligonucleotides. This protocol is applied to aluminum and copper oxide nanoparticles due to their great interest for the fabrication of highly energetic heterogeneous nanocomposites. The surface densities of streptavidin and biotinylated DNA oligonucleotides are precisely quantified combining atomic absorption spectroscopy with conventional dynamic light scattering and fluorometry and maximized to provide a basis for understanding the grafting mechanism. First, the streptavidin coverage is consistently below 20% of the total surface for both nanoparticles. Second, direct and unspecific grafting of DNA single strands onto Al and CuO nanoparticles largely dominates the overall functionalization process: ∼95% and 90% of all grafted DNA strands are chemisorbed on the CuO and Al nanoparticle surfaces, respectively. Measurements of hybridization efficiency indicate that only ∼5 and ∼10% of single-stranded oligonucleotides grafted onto the CuO and Al surfaces are involved in the hybridization process, corresponding precisely to the streptavidin coverage, as evidenced by the occupancy of 0.9 and 1.2 oligonucleotides per protein. The hybridization efficiency of single-stranded oligonucleotides chemisorbed on CuO and Al without streptavidin coating decreases to only ∼2%, justifying the use of streptavidin despite its poor surface occupancy. Finally, the structure of directly chemisorbed DNA strands onto oxide surfaces is examined and discussed.