PubReading [141] - Single-molecule fluorescence detection of a tricyclic nucleoside analogue - G. Samaan, B. Purse et al

Fluorescent nucleobase surrogates capable of Watson–Crick hydrogen bonding are essential probes of nucleic acid structure and dynamics, but their limited brightness and short absorption and emission wavelengths have rendered them unsuitable for single-molecule detection. Aiming to improve on these properties, we designed a new tricyclic pyrimidine nucleoside analogue with a push–pull conjugated system and synthesized it in seven sequential steps. The resulting C-linked 8-(diethylamino)benzo[b][1,8] naphthyridin-2(1H)-one nucleoside, which we name ABN, exhibits 3442 1⁄4 20 000 M1 cm1 and Fem,540 1⁄4 0.39 in water, increasing to Fem 1⁄4 0.50–0.53 when base paired with adenine in duplex DNA oligonucleotides. Single-molecule fluorescence measurements of ABN using both one-photon and two- photon exc ...