Organized arrays of individual DNA molecules tethered to supported lipid bilayers
Organized arrays of individual DNA molecules tethered to supported lipid bilayers
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DOI：
10.1021/la051944a
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发表时间：
2006-01-03
期刊：
影响因子：
3.9
通讯作者：
Greene, EC
中科院分区：
文献类型：
--
作者：
Granéli, A;Yeykal, CC;Greene, EC
许多单分子技术的一个未被认可的方面是，需要一个惰性表面，单个分子可以固定在这个表面上，而不会损害它们的生物完整性。在这里，我们提出了新的方法，将大的DNA分子连接到微流体样本室的表面，该样本室由于沉积了支撑的脂类双层而变得惰性。这些方法利用了两性离子脂提供的“生物友好”环境，但仍然允许DNA分子固定在表面的固定位置。我们还展示了一种新的方法来构建单个DNA分子的平行阵列，这些DNA分子在双层涂层的熔融二氧化硅表面上的特定位置组装。通过使用全内反射荧光显微镜来可视化阵列，可以在一个视场内同时监测数百个排列的DNA分子。这些分子阵列将允许并行处理多个单独的反应轨迹，从而显著提高基于荧光的单分子检测方法的吞吐能力。
An unappreciated aspect of many single-molecule techniques is the need for an inert surface to which individual molecules can be anchored without compromising their biological integrity. Here, we present new methods for tethering large DNA molecules to the surface of a microfluidic sample chamber that has been rendered inert by the deposition of a supported lipid bilayer. These methods take advantage of the "bio-friendly" environment provided by zwitterionic lipids, but still allow the DNA molecules to be anchored at fixed positions on the surface. We also demonstrate a new method for constructing parallel arrays of individual DNA molecules assembled at defined positions on a bilayer-coated, fused silica surface. By using total internal reflection fluorescence microscopy to visualize the arrays, it is possible to simultaneously monitor hundreds of aligned DNA molecules within a single field-of-view. These molecular arrays will significantly increase the throughput capacity of single-molecule, fluorescence-based detection methods by allowing parallel processing of multiple individual reaction trajectories.