I have two complementary oligonucleotides (about 25 nt). How long will they hybridize into dSDNA in a certain buffer at room temperature? Are there some differences between immobilization and fluid state?
Are you trying to base pair your two nucleotides. If yes, I usually do that by mixing them in a 1:1 molar ratio in PCR buffer (for Taq polymerase), heat them at 94 deg C for 3 minutes and then let the solution cool on the bench till it reaches room temperature. That should work.