DNA analysis by single molecule stretching in nanofluidic biochips
DNA analysis by single molecule stretching in nanofluidic biochips
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DOI：
10.1016/j.mee.2010.11.025
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发表时间：
2011-03-01
影响因子：
2.3
通讯作者：
Kristensen, A.
中科院分区：
文献类型：
--
作者：
Abad, E.;Juarros, A.;Kristensen, A.
在过去几年中，作为DNA分析工具，在纳米流体通道中通过限制拉伸单个DNA分子引起了极大的兴趣。我们已经根据使用高吞吐量纳米颗粒光刻（NIL）技术与硅底座和Pyrex盖的常规阳极粘结相结合，设计并制造了密封的微/纳米流体设备，用于DNA拉伸应用。使用此芯片，我们在台式荧光显微镜系统上进行了单分子成像。 Lambda噬菌体DNA用作模型样品来表征芯片。用荧光染料yoyo-1染色的Lambda-DNA的单分子在纳米渠道阵列中拉伸，并分析实验结果，以确定芯片中DNA的扩展因子和纳米渠内部直径的几何平均值。芯片的扩展比的确定提供了确定DNA大小的方法。这项工作的结果证明，开发的制造过程是制造单分子DNA生物芯片的一个很好的替代方法，它允许基于单分子DNA测序设备开发各种创新的生物/化学传感器。 （c）2010 Elsevier B.V.保留所有权利。
Stretching single DNA molecules by confinement in nanofluidic channels has attracted a great interest during the last few years as a DNA analysis tool. We have designed and fabricated a sealed micro/nanofluidic device for DNA stretching applications, based on the use of the high throughput NanoImprint Lithography (NIL) technology combined with a conventional anodic bonding of the silicon base and Pyrex cover. Using this chip, we have performed single molecule imaging on a bench-top fluorescent microscope system. Lambda phage DNA was used as a model sample to characterize the chip. Single molecules of lambda-DNA stained with the fluorescent dye YOYO-1 were stretched in the nanochannel array and the experimental results were analysed to determine the extension factor of the DNA in the chip and the geometrical average of the nanochannel inner diameter. The determination of the extension ratio of the chip provides a method to determining DNA size. The results of this work prove that the developed fabrication process is a good alternative for the fabrication of single molecule DNA biochips and it allows developing a variety of innovative bio/chemical sensors based on single-molecule DNA sequencing devices. (C) 2010 Elsevier B.V. All rights reserved.