首页> 美国卫生研究院文献>Nucleic Acids Research >Application of 2-cyanoethyl NNNN-tetraisopropylphosphorodiamidite for in situ preparation of deoxyribonucleoside phosphoramidites and their use in polymer-supported synthesis of oligodeoxyribonucleotides.
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Application of 2-cyanoethyl NNNN-tetraisopropylphosphorodiamidite for in situ preparation of deoxyribonucleoside phosphoramidites and their use in polymer-supported synthesis of oligodeoxyribonucleotides.

机译:2-氰基乙基NNNN-四异丙基亚磷酰胺在原位制备脱氧核糖核苷亚磷酰胺的应用及其在聚合物支持的寡聚脱氧核糖核苷酸合成中的应用。

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摘要

Deoxyribonucleoside phosphoramidites are prepared in situ from 5'-O,N-protected deoxyribonucleosides and 2-cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite with tetrazole as catalyst, and the solutions applied directly on an automatic solid-phase DNA synthesizer. Using LCAA-CPG support and a cycle time of 12.5 min, oligonucleotides of 16-25 bases are obtained with a DMT-efficiency per cycle of 98.0-99.3%. The crude and fully deblocked products are of a purity comparable to that obtained using purified phosphoramidites. In case of d(G)16 the product was difficult to analyse and a better product was not obtained using doubly protected (O-6 diphenylcarbamoyl) guanine.
机译:脱氧核糖核苷亚磷酰胺是由5'-O,N保护的脱氧核糖核苷和2-氰基乙基N,N,N',N'-四异丙基亚磷酰胺以四唑为催化剂原位制备的,溶液直接应用于自动固相DNA合成仪上。使用LCAA-CPG支持和12.5分钟的循环时间,可获得16-25个碱基的寡核苷酸,每个循环的DMT效率为98.0-99.3%。粗产物和完全解嵌段的产物的纯度与使用纯化的亚磷酰胺获得的纯度相当。在d(G)16的情况下,该产品难以分析,并且使用双重保护的(O-6二苯基氨基甲酰基)鸟嘌呤无法获得更好的产品。

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