Visual Detection of Labeled Oligonucleotides Using Visible-Light-Polymerization-Based Amplification

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• 作者：Ryan R. Hansen ; Hadley D. Sikes ; Christopher N. Bowman
• 刊名：Biomacromolecules
• 出版年：2008
• 出版时间：January 2008
• 年：2008
• 卷：9
• 期：1
• 页码：355 - 362
• 全文大小：934K
• 年卷期：v.9,no.1(January 2008)
• ISSN：1526-4602

文摘

DNA biochip technology holds potential for highly parallel, rapid, and sensitive genetic diagnostic screening of target pathogens and disease biomarkers. A primary limitation involves a simultaneous, sequence-specific identification of low copy number target polynucleotides using a clinically appropriate detection methodology that implements only inexpensive detection instrumentation. Here, a rapid (20 min), nonenzymatic method of signal amplification utilizing surface-initiated photopolymerization is presented in glass microarray format. Visible light photoinitiators covalently coupled to streptavidin were used to bind biotin-labeled capture sequences. Amplification was achieved through subsequent contact with a monomer solution and the appropriate light exposure to generate 20−240-nm-thick hydrogel layers exclusively from spots containing the biotin-labeled DNA. An amplification factor of 10⁶ to 10⁷ was observed as well as a detectable response generated from as low as ~10⁴ labeled oligonucleotides using minimal instrumentation, such as an optical microscope or CCD camera.