RESUMO
Facile modification of oligodeoxyribonucleotides is required for efficient immobilization to a pre-activated glass surface. This report presents an oligodeoxyribonucleotide which contains a hairpin stem-loop structure with multiple phosphorothioate moieties in the loop. These moieties are used to anchor the oligo to glass slides that are pre-activated with bromoacetamidopropylsilane. The efficiency of the attachment reaction was improved by increasing the number of phosphorothioates in the loop, as shown in the remarkable enhancement of template hybridization and single base extension through catalysis by DNA polymerase. The loop and stem presumably serve as lateral spacers between neighboring oligodeoxyribonucleotides and as a linker arm between the glass surface and the single-stranded sequence of interest. The oligodeoxyribonucleotides of this hairpin stem-loop architecture with multiple phosphorothioate moieties have broad application in DNA chip-based gene analysis.
Assuntos
Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Sequência de Bases , Primers do DNA/química , Primers do DNA/genética , Primers do DNA/metabolismo , DNA Polimerase Dirigida por DNA/metabolismo , Vidro , Estrutura Molecular , Conformação de Ácido Nucleico , Hibridização de Ácido Nucleico , Oligodesoxirribonucleotídeos/genética , Silanos , Moldes GenéticosRESUMO
Tetrakis(triphenylphosphine)palladium(0) effects the regio- and stereo-selective alkylation of 2-acetoxy-5,6-dihydro-2H-pyrans and 1-S-acetyl-1-thiohex-2-enopyranosides. Use of stabilized carbanions resulted in the formation of alkylated dihydropyrans with net retention at the oxygen-bearing carbon atom. Examples include the preparation of 2-[acetamidobis(ethoxycarbonyl)methyl]-5,6-dihydro-2H-pyran, 2-[acetamidobis(methoxycarbonyl)methyl]-6-ethoxy-5,6-dihydro-2H-py ran, and 2-[acetamidobis(methoxycarbonyl)methyl]-6-methoxymethyl-5,6-dihydr o-2H- pyran. Use of nonstabilized carbanions, such as arylzinc chlorides, resulted in the formation of alkylated dihydropyrans and C-glycosyl compounds with net inversion at the oxygen-bearing carbon atom. Examples include the preparation of 2-[(6-ethoxy-5,6-dihydro-2H-pyranyl)methyl]-4,4-dimethyl-2-oxazoli ne, trans-methoxymethyl-2-phenyl-5,6-dihydro-2H-pyran, trans-methoxymethyl-2-vinyl-5,6-dihydro-2H-pyran, trans-2-[2,2-bis(ethoxy)ethyl]-6-methoxymethyl-5,6-dihydro-2H-pyran, (4,6-di-O-methyl-2,3-dideoxy-alpha-D-erythro-hex-2-enopyranosyl)be nzene, (2,3-dideoxy-4,6-di-O-methyl-beta-D-erythro-hex-2-enopyranosyl)ben zene, 1-(2,3-dideoxy-4,6-di-O-methyl-alpha and -beta-D-erythro-hex-2-enopyranosyl)naphthalene, 4-(2,3-dideoxy-4,6-di-O-methyl-alpha- and -beta-D-erythro-hex-2-enopyranosyl)toluene, and 1-(2,3-trideoxy-4-O-methyl-alpha-L-erythro-hex-2-enopyranosyl)naph thalene.
