McCormick Laurette C, Slater Gary W
University of Ottawa, Ottawa, Ontario, Canada.
Electrophoresis. 2007 Feb;28(4):674-82. doi: 10.1002/elps.200600590.
Recent advancements in DNA sequencing by end-labeled free-solution electrophoresis (ELFSE) show the promise of this novel technique which overcomes the need for a gel by using a label (or drag-tag) to render the free solution mobility of the DNA size-dependent. It is the attachment of an uncharged drag-tag molecule of a set size to various lengths of DNA in the sample that selectively slows down smaller DNA chains which have less force to pull the drag-tag than larger DNA. So far, only globally random coil conformations have been associated with ELFSE, i.e., the DNA and the label together form a single, undeformed hydrodynamic unit. This paper investigates the conditions under which the DNA and label will segregate into two hydrodynamically distinct units, based on a theoretical approach developed for the electrophoresis of polyampholytes. Optimal experimental conditions tailored to the available label sizes and voltages are predicted along with insight into ideal label architecture.
通过末端标记自由溶液电泳（ELFSE）进行DNA测序的最新进展显示了这种新技术的前景，该技术通过使用标签（或拖尾标签）使DNA在自由溶液中的迁移率取决于其大小，从而无需凝胶。将特定大小的不带电荷的拖尾标签分子连接到样品中不同长度的DNA上，会选择性地减缓较小的DNA链，因为它们拉动拖尾标签的力比大的DNA链小。到目前为止，ELFSE仅与整体随机卷曲构象相关，即DNA和标签一起形成单个未变形的流体动力学单元。本文基于为聚两性电解质电泳开发的理论方法，研究了DNA和标签将分离成两个流体动力学上不同单元的条件。预测了针对可用标签大小和电压量身定制的最佳实验条件，并深入了解了理想的标签结构。
