Sarac Ivo, Meier Chris
Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
Chemistry. 2015 Nov 9;21(46):16421-6. doi: 10.1002/chem.201502844. Epub 2015 Aug 25.
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.
报道了一种快速、高产且可靠的合成DNA和RNA 5'-三磷酸酯的方法。通过自动寡核苷酸合成法合成DNA或RNA寡核苷酸后，将5-氯-亚水杨基-N,N-二异丙基亚磷酰胺偶联到5'-末端。使用标准寡核苷酸合成中使用的相同氧化试剂氧化形成的5'-亚磷酸酯，得到5'-环Sal-寡核苷酸。负载型5'-环Sal-寡核苷酸与焦磷酸反应，生成相应的5'-三磷酸酯。从载体上切割后，以高纯度和优异产率获得5'-三磷酸化的DNA和RNA寡核苷酸。整个反应序列适用于标准寡核苷酸合成仪。
