Electrophoresis of end-labeled DNA: Theory and experiment
Electrophoresis of end-labeled DNA: Theory and experiment
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DOI：
10.1103/physreve.81.031918
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发表时间：
2010-03-01
影响因子：
2.4
通讯作者：
Archer, Lynden A.
中科院分区：
文献类型：
--
作者：
Lau, Henry W.;Archer, Lynden A.
用一个简单的哑铃模型研究了末端标记DNA在自由溶液电泳过程中的动力学行为。我们研究了外加电场、标签尺寸和链刚度对DNA构象和电泳迁移率的影响。高的外加电场被预测放大的尺寸依赖性的迁移率，并产生一个非单调依赖的电泳迁移率上施加的电场。利用标签大小和DNA链刚度提高分辨率的有效性也讨论了DNA变形的背景下。为了评估最突出的模型预测，我们使用毛细管电泳实验来表征的大小和字段依赖性的流动性的dsDNA片段（300 bp-2 kbp）的末端功能化与链霉亲和素。我们的实验结果被发现是在一般良好的雅阁与期望的基础上的哑铃模型。我们讨论了这些研究结果的影响，快速，基于尺寸的DNA在自由溶液中的分离。
The dynamic behavior of end-labeled DNA during free-solution electrophoresis is investigated using a simple dumbbell model for the labeled DNA. We study the effect of the applied field, label size, and chain stiffness on DNA conformation and electrophoretic mobility. High applied fields are predicted to magnify the size-dependence of mobility and to yield a nonmonotonic dependence of electrophoretic mobility on applied field. The effectiveness of leveraging label size and DNA chain stiffness for improving resolution is also discussed in the context of DNA deformation. To evaluate the most salient model predictions, we use capillary electrophoresis experiments to characterize the size-and field-dependent mobility of dsDNA fragments (300 bp-2 kbp) end-functionalized with streptavidin. Our experimental results are found to be in generally good accord with expectations based on the dumb-bell model. We discuss implications of these findings for fast, size-based separation of DNA in free solution.