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纳米流体通道的制造及DNA分子的操控

Nanofluidic channel fabrication and manipulation of DNA molecules.

作者信息

Wang Kai-Ge, Niu Hanben

机构信息

Institute of Photonics and Photonic Technology, Northwest University, Shaanxi, China.

出版信息

Methods Mol Biol. 2009;544:17-27. doi: 10.1007/978-1-59745-483-4_2.

Abstract

Confining DNA molecules in a nanofluidic channel, particularly in channels with cross sections comparable to the persistence length of the DNA molecule (about 50 nm), allows the discovery of new biophysical phenomena. This sub-100 nm nanofluidic channel can be used as a novel platform to study and analyze the static as well as the dynamic properties of single DNA molecules, and can be integrated into a biochip to investigate the interactions between protein and DNA molecules. For instance, nanofluidic channel arrays that have widths of approximately 40 nm, depths of 60 nm, and lengths of 50 mum are created rapidly and exactly by a focused-ion beam milling instrument on a silicon nitride film; and the open channels are sealed with anodic bonding technology. Subsequently, lambda phage DNA (lambda-DNA; stained with the fluorescent dye, YOYO-1) molecules are introduced into these nanoconduits by capillary force. The movements of the DNA molecules, e.g. stretching, recoiling, and transporting along channels, are studied with fluorescence microscopy.

摘要

将DNA分子限制在纳米流体通道中,特别是在横截面与DNA分子的持久长度(约50纳米)相当的通道中,可以发现新的生物物理现象。这种亚100纳米的纳米流体通道可作为一个新颖的平台,用于研究和分析单个DNA分子的静态和动态特性,并且可以集成到生物芯片中以研究蛋白质与DNA分子之间的相互作用。例如,通过聚焦离子束铣削仪器在氮化硅膜上快速精确地制造出宽度约为40纳米、深度为60纳米、长度为50微米的纳米流体通道阵列;并用阳极键合技术密封开放通道。随后,通过毛细作用力将λ噬菌体DNA(λ-DNA;用荧光染料YOYO-1染色)分子引入这些纳米管道中。利用荧光显微镜研究DNA分子的运动,如拉伸、回缩以及沿通道的运输。

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