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基于寡核苷酸固定在经活性丙烯酸-丙烯酰胺共聚物修饰的玻璃表面的寡核苷酸微阵列。

Oligo-microarray based on oligonucleotide immobilization on glass surface modified with activated acrylic acid-co-acrylamide copolymer.

作者信息

Wu Qinghua, Ma Wenli, Zhu Lina, Guo Qiuye, Zhang Bao, Kulageri Shivappa M, Zheng Wenling

机构信息

Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China.

出版信息

J Biomed Mater Res B Appl Biomater. 2008 Oct;87(1):67-72. doi: 10.1002/jbm.b.31069.

Abstract

Oligo-microarray can be fabricated on modified surface of glass slides involving immobilization of oligonucleotides. In this study, glass slides were modified with acrylic acid-co-acrylamide copolymer and EDC (1-ethyl-3(3-dimethylaminopropyl)-carbodiimide)/NHS (N-hydroxysuccinimide) by covalent linkage method, and covalent immobilization of unmodified oligonucleotide on the glass surface was achieved. The platform with binding of stable and sensitive oligonucleotides was prepared for oligo-microarray fabrication. An optimum concentration of 0.5 g/L was used for spotting. The lengths of 26 to 70 mer oligonucleotides were immobilized on the surface efficiently. The spots were approximately 160 nm in diameter and their mean value of fluorescent intensity was measured in the range of 3.2 x 10(4) to 7.3 x 10(4) after hybridization. The modified glass surface was scanned by SEM and the dendritic structure was observed. Results showed that the process of preparation of the modified glass surface was simple and cost effective, and the modified surface can be used for the oligo-microarray fabrication and also attachment of protein, PNA etc.

摘要

寡核苷酸微阵列可以在经过修饰的载玻片表面制备,这涉及到寡核苷酸的固定化。在本研究中,通过共价连接法用丙烯酸 - 丙烯酰胺共聚物以及1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS)对载玻片进行修饰,实现了未修饰寡核苷酸在玻璃表面的共价固定化。为制备寡核苷酸微阵列搭建了结合稳定且灵敏的寡核苷酸的平台。点样采用0.5 g/L的最佳浓度。长度为26至70个碱基的寡核苷酸被有效地固定在表面。杂交后斑点直径约为160纳米,其荧光强度平均值在3.2×10⁴至7.3×10⁴范围内。通过扫描电子显微镜(SEM)对修饰后的玻璃表面进行扫描,观察到树枝状结构。结果表明,修饰玻璃表面的制备过程简单且成本效益高,该修饰表面可用于寡核苷酸微阵列的制备以及蛋白质、肽核酸等的附着。

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