10.1021/jo970872s.s001 Michael C. Pirrung Michael C. Pirrung Lara Fallon Lara Fallon Glenn McGall Glenn McGall Proofing of Photolithographic DNA Synthesis with 3‘,5‘-Dimethoxybenzoinyloxycarbonyl-Protected Deoxynucleoside Phosphoramidites American Chemical Society 1998 probe GTAGCATCTT hybridization nonpolar medium homopolymers target deoxyeicosanucleotide Proofing irradiation sequence method solvent efficiency amidite monomer Photolithographic DNA Synthesis nucleoside building blocks phosphoramidite cycle yields fluorescein pyrimidine array purine Phosphoramidite synthesis microchip format hybridized covalently photolabile al Photolysis rates nm Deprotection McGall GAC derivatized glass surfaces data DMBOC AAXTAXCTAC Deoxynucleoside 16 decanucleotides Surface fluorescence imaging 1998-01-01 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/Proofing_of_Photolithographic_DNA_Synthesis_with_3_5_-Dimethoxybenzoinyloxycarbonyl-Protected_Deoxynucleoside_Phosphoramidites/3707013 We have evaluated in a microchip format the photochemical solid-phase phosphoramidite DNA synthesis method we previously developed. A set of nucleoside building blocks with “easy-off” base protecting groups was prepared bearing photolabile 5‘-O-dimethoxybenzoincarbonate (DMBOC) groups. Photolysis rates and cycle yields for these DMBOC-protected nucleotides covalently attached to planar, derivatized glass surfaces were determined by fluorescence imaging-based methods earlier developed by McGall et al. and described in detail elsewhere. Data were obtained for both 280/310 and 365/400 nm irradiation in a range of solvents. Deprotection of the DMBOC occurs fastest in a nonpolar medium or without solvent. The coupling efficiency of these amidites in the synthesis of homopolymers was determined to be in the range 80−97%, with purines generally showing lower efficiency than pyrimidines. These DMBOC-protected monomers were used to prepare a 4 × 4 array of 16 decanucleotides of the sequence 5‘-AAXTAXCTAC−chip, where X = A, C, G, or T. The array was hybridized with a target deoxyeicosanucleotide of the sequence fluorescein-5‘-CTGAACG<b>GTAGCATCTT</b>GAC. Surface fluorescence imaging demonstrated sequence-specific hybridization to this probe.