高等学校化学学报

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (6): 1128.

• 研究快报 • 上一篇    下一篇

玻璃微流控通道中水凝胶固定寡核苷酸探针的方法及应用

张宇1,2, 于浩1, 董秀玲3, 秦建华1, 林炳承1   

  1. 1. 中国科学院大连化学物理研究所, 大连 116023;
    2. 中国科学院研究生院, 北京 100049;
    3. 大连市第三人民医院, 大连 116033
  • 收稿日期:2009-02-16 出版日期:2009-06-10 发布日期:2009-06-10
  • 通讯作者: 林炳承, 男, 研究员, 主要从事微流控芯片的研究, E-mail: bclin@dicp.ac.cn; 秦建华, 女, 研究员, 主要从事微流控芯片的研究, E-mail: jhqin@dicp.ac.cn
  • 基金资助:

    国家自然科学基金(批准号: 20635030, 20575067)和科技部“八六三”计划(批准号: 2006AA02Z305)资助.

Immobilization of DNA Hydrogel Plugs in Glass Microfluidic Channels and Its Applications

ZHANG Yu1,2, YU Hao1, DONG Xiu-Ling3, QIN Jian-Hua1*, LIN Bin-Cheng1*   

  1. 1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;
    2. Graduate School of Chinese Academy of Sciences, Beijing 100049, China;
    3. Dalian Third Municipal Hospital, Dalian 116033, China
  • Received:2009-02-16 Online:2009-06-10 Published:2009-06-10
  • Contact: QIN Jian-Hua, E-mail: bclin@dicp.ac.cn; LIN Bin-Cheng, E-mail: jhqin@dicp.ac.cn
  • Supported by:

    国家自然科学基金(批准号: 20635030, 20575067)和科技部“八六三”计划(批准号: 2006AA02Z305)资助.

PDF (PC)

被引次数

11

摘要：

本文报道在玻璃微流控通道中, 利用光聚合的方法, 将丙烯基修饰的单链寡核苷酸探针固定在丙烯酰胺水凝胶中. 通道采用亲和硅烷修饰, 聚合后的水凝胶共价锚定在硅烷化的通道壁上. 该水凝胶结构稳定, 不受温度和pH值的影响. 在电泳条件下水凝胶为多孔基质, 可以通过杂交反应捕获样品中的特异性靶序列. 同一块芯片经变性及重新杂交, 可重复使用多次. 由于电场有增强杂交的作用, 因此样品分析时间可由以往的几小时甚至十几小时缩短至5 min.

关键词: 微流控芯片, 丙烯酰胺水凝胶, DNA探针固定, 核酸杂交

Abstract:

A method of DNA probes immobilization and specific target sequences capture in a microfluidic chip was presented. Acrydite-modified DNA probes are immobilized in silanized glass microfluidic channels via photopolymerization in a polyacrylamide matrix. The resulting polymeric, hydrogel plugs are porous under electrophoretic conditions, and the immobilized DNA probes can be hybridized with fluorescence labeled complementary DNA. The total analysis process can be completed within 5 min, and the limit of detection is 0.1 μmol/L. This method is simple, rapid and feasible. The double-stranded DNA can be chemically denatured, and the chip is reusable. The conditions for photopolymerization, hybridization, and denaturation were discussed as well.

Key words: Microfluidic chip, Acrylamide hydrogel, DNA probes immobilization, Hybridization

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