For the first time, the covalent immobilization of oligonucleotides double helices onto surfaces prior to sequential denaturation and rehybridization is proven to lead to optimal hybridization efficiency. Two indirect methods were used for monitoring these reactions in situ: the quartz crystal microbalance with dissipation monitoring (QCM-D) and the wavelength interrogated optical sensor (WIOS, Bright Reader). Both techniques led to the result that with this immobilization approach one could reach nearly 100% hybridization efficiency. Moreover, applying the polymer theory to the adsorption of nucleotide sequences onto surfaces, we demonstrate, that for single stranded sequences the coil conformation prevails over the stretch one.