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高效液相色谱法纯化含有碱基和骨架修饰序列的合成寡脱氧核糖核苷酸。

HPLC purification of synthetic oligodeoxyribonucleotides containing base- and backbone-modified sequences.

作者信息

Green A P, Burzynski J, Helveston N M, Prior G M, Wunner W H, Thompson J A

机构信息

Puresyn, Inc., Malvern, PA, USA.

出版信息

Biotechniques. 1995 Nov;19(5):836-41.

PMID:8588925
Abstract

Applications for a new polymer resin, PolyFlo, are described for both the small-scale and large-scale purification of synthetic oligodeoxyribonucleotides varying in length from 18-41 bases. The unique properties of this innovative resin provide > 95% purified full-length oligodeoxyribonucleotides with greater than 90% yield starting from either crude trityl-on or trityl-off unmodified as well as base (biotin)- or backbone (e.g., phosphorothioate)-modified products. Full biological activity of recovered nucleic acid is retained, and the resin is capable of removing contaminating endotoxins during purification. The resin performance is predictable and reliable. The resin can be regenerated easily and is particularly economic when employed directly in ammonia or with the trityl-off option. PolyFlo meets the requirements of current Good Manufacturing Practices.

摘要

一种新型聚合物树脂PolyFlo的应用被描述用于小规模和大规模纯化长度在18至41个碱基之间变化的合成寡脱氧核糖核苷酸。这种创新树脂的独特性能可从粗制的三苯甲基保护型或三苯甲基脱除型未修饰以及碱基(生物素)或主链(例如硫代磷酸酯)修饰的产物中提供大于95%纯化的全长寡脱氧核糖核苷酸,产率大于90%。回收的核酸保留了完全的生物活性,并且该树脂在纯化过程中能够去除污染的内毒素。树脂性能可预测且可靠。该树脂易于再生,并且直接用于氨中或采用三苯甲基脱除选项时特别经济。PolyFlo符合现行良好生产规范的要求。

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