Department of Chemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada.
Biomicrofluidics. 2010 Sep 30;4(3):32209. doi: 10.1063/1.3463720.
Two simple gold nanoparticle (GNP)-based DNA analysis methods using a microfluidic device are presented. In the first method, probe DNA molecules are immobilized on the surface of a self-assembled submonolayer of GNPs. The hybridization efficiency of the target oligonulceotides was improved due to nanoscale spacing between probe molecules. In the second method, target DNA molecules, oligonulceotides or polymerase chain reaction (PCR) amplicons, are first bound to GNPs and then hybridized to the immobilized probe DNA on a glass slide. With the aid of GNPs, we have successfully discriminated, at room temperature, between two PCR amplicons (derived from closely related fungal pathogens, Botrytis cinerea and Botrytis squamosa) with one base-pair difference. DNA analysis on the microfluidic chip avoids the use of large sample volumes, and only a small amount of oligonucelotides (8 fmol) or PCR products (3 ng), was needed in the experiment. The whole procedure was accomplished at room temperature in 1 h, and apparatus for high temperature stringency was not required.
介绍了两种基于金纳米粒子（GNP）的微流控芯片上的简单 DNA 分析方法。在第一种方法中，探针 DNA 分子被固定在自组装的 GNP 亚单层表面上。由于探针分子之间的纳米级间距，目标寡核苷酸的杂交效率得到了提高。在第二种方法中，首先将靶 DNA 分子、寡核苷酸或聚合酶链反应（PCR）扩增子与 GNP 结合，然后与玻璃载玻片上固定的探针 DNA 杂交。借助 GNP，我们成功地在室温下区分了两个 PCR 扩增子（源自密切相关的真菌病原体 Botrytis cinerea 和 Botrytis squamosa），它们只有一个碱基差异。微流控芯片上的 DNA 分析避免了使用大体积的样品，实验中只需要少量的寡核苷酸（8 fmol）或 PCR 产物（3 ng）。整个过程在 1 小时内于室温下完成，不需要高温严格性的仪器。
