We investigated the molecular recognition between the amphiphile AzoAde, which is composed of azobenzene in the hydrophobic and adenine in the hydrophilic portion of the molecule, and oligonucleotides having a homogeneous base (dA(30), dT(30), dG(30), and dC(30)) at the air-water interface. On the basis of the complementary base-pairing of DNA in the duplex, orderly arrangement of AzoAde on templated dT(30) was examined using pi-A isotherm, UV-vis RAS, FT-IR RAS, and XPS measurements. Although there was little interaction between AzoAde and mismatched oligonucleotides (dA(30), dG(30) and dC(30)), AzoAde prepared on a dT(30) interacted with dT(30), subsequently forming a J-form assembly at the air-water interface. AFM observation of the LB films revealed the nanostructure of the J-formed AzoAde monolayer on the dT(30) subphase as well as the domain structures of the H-formed monolayers on the other oligonucleotide subphases. Therefore, dT(30) has a potential application as a template for assembling AzoAde at the air-water interface.