title: Efficient Automated Solid-Phase Synthesis of DNA and RNA 5'-Triphosphates.
abstract: A fast, high-yielding and reliable method for the synthesis of DNA- and RNA 5'-triphosphates is reported. After synthesizing DNA or RNA oligonucleotides by automated oligonucleotide synthesis, 5-chloro-saligenyl-N,N-diisopropylphosphoramidite was coupled to the 5'-end. Oxidation of the formed 5'-phosphite using the same oxidizing reagent used in standard oligonucleotide synthesis led to 5'-cycloSal-oligonucleotides. Reaction of the support-bonded 5'-cycloSal-oligonucleotide with pyrophosphate yielded the corresponding 5'-triphosphates. The 5'-triphosphorylated DNA and RNA oligonucleotides were obtained after cleavage from the support in high purity and excellent yields. The whole reaction sequence was adapted to be used on a standard oligonucleotide synthesizer.
authors.affiliation: Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
authors.forename: Ivo
authors.initials: I
authors.lastname: Sarac
authors.affiliation: Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany). chris.meier@chemie.uni-hamburg.de.
authors.forename: Chris
authors.initials: C
authors.lastname: Meier
mesh_terms: D001592:Benzyl Alcohols / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry*; D011122:Polyphosphates; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry; D060327:Solid-Phase Synthesis Techniques
chemical_list: C000608191:5-chloro-saligenyl-N,N-diisopropylphosphoramidite; D001592:Benzyl Alcohols; D007202:Indicators and Reagents; D009841:Oligonucleotides; D009943:Organophosphorus Compounds; D011122:Polyphosphates; D012313:RNA; D004247:DNA; C005692:triphosphoric acid
keywords: 5′-triphosphates; cycloSal; oligonucleotides; phosphoramidite chemistry; solid-phase synthesis