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In this study, we have conducted a high-resolution nuclear magnetic resonance (NMR) spectroscopic research on option MDB-forming sequences and found that an MDB may be created stably with a purine within the small groove, which has never been noticed in any formerly reported DNA type II tetraloops. Our refined NMR option structures of the two MDBs created by 5'-CTTG CATG-3' and 5'-CTTG CGTG-3' expose that the 6th purine residue was driven into the minor groove via base-base stacking using the se