Electrostretching DNA molecules using polymer-enhanced media within microfabricated devices
Electrostretching DNA molecules using polymer-enhanced media within microfabricated devices
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DOI：
10.1021/ac025551h
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发表时间：
2002-07-15
影响因子：
7.4
通讯作者：
Burns, MA
中科院分区：
文献类型：
--
作者：
Namasivayam, V;Larson, RG;Burns, MA
在本文中，我们证明了固定和拉伸的单噬菌体DNA分子在微流体系统中使用交流领域。我们提出了一种新的“巯基-金”为基础的固定化技术，用于固定一个特定的结束（T端）的DNA分子到金电极。聚合物增强介质（类似于Tris-HCl中的3.75重量%线性聚丙烯酰胺）用于获得荧光染色的DNA分子的完全拉伸构型（21 μ m）。我们还提出了一个优化的微电极设计与点电极和电极间距为20 μ m的拉伸DNA分子与交流电场（1 MHz，3 × 10（5）V/m）。最后，使用这些技术，我们将单个DNA分子固定在一个电极边缘，拉伸分子，并将另一端固定在相邻的电极边缘，在微加工设备内的两个电极之间形成桥梁。
In this paper, we demonstrate immobilization and stretching of single lambda-phage DNA molecules within microfluidic systems using ac fields. We present a novel "thiol-on-gold"-based immobilization technique for fixing one specific end (T end) of a DNA molecule onto a gold electrode. A polymer-enhanced medium (similar to3.75 wt % linear polyacrylamide in Tris-HCl) is used to obtain fully stretched configurations (21 mum) of fluorescently stained lambda-DNA molecules. We also present an optimized microelectrode design with pointed electrodes and an electrode spacing of 20 mum for stretching DNA molecules with an ac field (1 MHz, 3 x 10(5) V/m). Finally, using these techniques, we immobilize a single DNA molecule at one electrode Edge, stretch the molecule, and fix the other end at an adjacent electrode edge, forming a bridge between two electrodes within a microfabricated device.