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[一种将寡核苷酸固定在含羧基尼龙膜上用于核酸杂交问题的新方法]

[A new approach to the problem of immobilizing oligonucleotides on carboxyl-containing nylon membranes for nucleic acid hybridization].

作者信息

Ivanovskaia M G, Kozlov I A, Naryshkin N A, Shabarova Z A

出版信息

Bioorg Khim. 1995 Jul;21(7):535-8.

PMID:7488269
Abstract

A simple and efficient method of the covalent immobilization of oligonucleotides on carboxyl-containing nylon membranes was proposed. The method is based on the reaction between a postsynthetically introduced aminoalkyl group of an oligonucleotide and membrane carboxyl catalyzed by a water soluble N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide. Judging from the mechanism of carboxyl group activation by the carbodiimide, optimal conditions of the immobilization were selected, which made it possible to increase the immobilization efficiency and carrier capacity as compared with those described previously. Using a 23-unit membrane-immobilized oligonucleotide as an example, the oligonucleotides immobilized were shown to be capable of hybridizing with complementary sequences.

摘要

提出了一种将寡核苷酸共价固定在含羧基尼龙膜上的简单有效方法。该方法基于水溶性N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺催化的寡核苷酸合成后引入的氨基烷基与膜羧基之间的反应。从碳二亚胺活化羧基的机理出发,选择了最佳固定条件,与先前描述的条件相比,提高了固定效率和载体容量。以一个23个单元的膜固定寡核苷酸为例,证明固定的寡核苷酸能够与互补序列杂交。

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