The molecular structures of polyamide barrier layers in reverse osmosis membranes, made by interfacial polymerization of m-phenylenediamine and trimesoyl chloride under different reaction and post-treatment conditions, were characterized by grazing incidence wide-angle X-ray scattering (GIWAXS). The molecular backbone packing is consistent with two different aromatic molecular packing motifs (parallel and perpendicular) with preferential surface-induced orientation. The results suggest that the perpendicular, T-shaped, packing motif (5 Å spacing) might be associated with optimal membrane permeance, compared with the parallel packings (3.5-4.0 Å spacings).