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通过亚微米圆柱玻璃毛细管基质快速筛选 DNA。

Fast DNA sieving through submicrometer cylindrical glass capillary matrix.

机构信息

Department of Electronic and Computer Engineering, Hong Kong University of Science and Technology , Clear Water Bay, Kowloon, Hong Kong.

出版信息

Anal Chem. 2014 Jan 7;86(1):737-43. doi: 10.1021/ac4031994. Epub 2013 Dec 12.

Abstract

Here, we report on DNA electrophoresis through a novel artificial sieving matrix based on the highly regular submicrometer cylindrical glass capillary segments alternatingly arranged with wells. Such round capillaries pose a higher-order confinement resulting in a lower partition coefficient and greater entropic energy barrier while limiting the driving field strength to a small fraction of the applied electric field. In return, the separation can be performed at high average field strengths (up to 1.6 kV/cm) without encountering the field-dependent loss of resolving power. This leads to fast DNA sieving as demonstrated here on the capillaries of 750 nm in diameter. The 600 bp to 21 kbp long chains are shown to resolve within 4 min after having undergone a fairly limited number of entropic barriers (128 in total). The capillary matrix also exhibits a critical field threshold below which DNA bands fail to launch, and this occurs at a considerably greater magnitude than in other matrixes. The submicrometer capillaries are batch-fabricated on silicon through a fabrication process that does not require high-resolution advanced lithography or well-controlled wafer bonding techniques to define their critical dimension.

摘要

在这里,我们报告了一种基于高度规则的亚微米级圆柱形玻璃毛细管段与井交替排列的新型人工筛分基质的 DNA 电泳。这种圆形毛细管构成了更高阶的限制,导致较低的分配系数和更大的熵能量障碍,同时将驱动场强限制在施加电场的一小部分。作为回报,分离可以在较高的平均场强下进行(高达 1.6 kV/cm),而不会遇到分辨率随场变化的损失。这导致快速的 DNA 筛分,如这里在直径为 750nm 的毛细管上所示。600bp 到 21 kbp 长的链在经过相当有限数量的熵障碍(总共 128 个)后,在 4 分钟内得到解决。毛细管基质还表现出一个临界场阈值,低于该阈值时 DNA 带无法启动,而这一阈值比其他基质要大得多。亚微米级毛细管是通过一种制造工艺在硅片上批量制造的,这种工艺不需要高分辨率的先进光刻技术或精密控制的晶圆键合技术来定义其关键尺寸。

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