通过可更换毛细管凝胶电泳对寡脱氧核苷酸及其硫代磷酸酯修饰进行单碱基分辨率分析。

Single-based resolution for oligodeoxynucleotides and their phosphorothioate modifications by replaceable capillary gel electrophoresis.

作者信息

Chen Rong, Luo Xuefang, Di Xin, Li Ying, Sun Yuqing, Hu Yuzhu

机构信息

Department of Analytical Chemistry, China Pharmaceutical University, Box 41, 24 Tongjia Lane, Nanjing 210009, PR China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Nov 7;843(2):334-8. doi: 10.1016/j.jchromb.2006.05.034. Epub 2006 Jun 30.

DOI:10.1016/j.jchromb.2006.05.034

PMID:16809077

Abstract

A replaceable capillary gel electrophoretic (replaceable CGE) method was developed for the separation of two sets of model compounds of single-stranded oligodeoxynucleotide mixtures (18-20 mers), phosphodiester oligodeoxynucleotides (PO-ODNs) and their phosphorothioate modifications (PS-ODNs), with equal sequences differing in a single base. Polyethylene glycol (PEG) 35000 was chosen as the sieving matrix. It was confirmed that PEG polymer solution less influenced resolutions of the PS-ODNs compared with those of the PO-ODNs, while acetonitrile used as an additive in the system improved the separation significantly. It was also noticed that the effect of temperature on separation was much larger than that of denaturant urea.

摘要

开发了一种可替换毛细管凝胶电泳（可替换CGE）方法，用于分离两组单链寡脱氧核苷酸混合物（18 - 20聚体）的模型化合物，即磷酸二酯寡脱氧核苷酸（PO - ODNs）及其硫代磷酸酯修饰物（PS - ODNs），它们具有相同序列，仅单个碱基不同。选择聚乙二醇（PEG）35000作为筛分基质。已证实，与PO - ODNs相比，PEG聚合物溶液对PS - ODNs分辨率的影响较小，而系统中用作添加剂的乙腈显著改善了分离效果。还注意到温度对分离的影响远大于变性剂尿素的影响。