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Self-Associating Block Copolymer Networks for Microchip Electrophoresis Provide Enhanced DNA Separation via 'Inchworm' Chain Dynamics

机译:用于微芯片电泳的自缔合嵌段共聚物网络通过““”链动力学提供增强的DNA分离

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摘要

We describe a novel class of DNA separation media for microchip electrophoresis, "physically cross-linked" block copolymer networks, which provide rapid (<4.5 min) and remarkably enhanced resolution of DNA in a size range critical for genotyping. Linear poly(acrylamide-co-dihexy-lacrylamide) (LPA-co-DHA) comprising as little as 0.13 mol(percent) dihexylacrylamide yields substantially improved electrophoretic DNA separations compared to matched molar mass linear polyacrylamide. Single-molecule videomicroscopic images of DNA electrophoresis reveal novel chain dynamics in LPA-co-DHA matrixes, resembling inchworm movement, to which we attribute the increased DNA resolution. Substantial improvements in DNA peak separation are obtained, in particular, in LPA-co-DHA solutions at polymer/copolymer concentrations near the interchain entanglement threshold. Higher polymer concentrations yield enhanced separations only for small DNA molecules (<120 base pairs). Hydrophobically cross-linked networks offer advantages over conventional linear polymers based on enhanced separation performance (or speed) and over chemically cross-linked gels because hydrophobic cross-links can be reversibly broken, allowing facile microchannel loading.
机译:我们描述了一种用于微芯片电泳的新型“ DNA分离介质”,即“物理交联的”嵌段共聚物网络,该网络可在对基因分型至关重要的大小范围内提供快速(<4.5分钟)和显着增强的DNA分辨率。与匹配摩尔质量的线性聚丙烯酰胺相比,包含低至0.13 mol(%)二己基丙烯酰胺的线性聚(丙烯酰胺-共-二己基-丙烯酰胺)(LPA-co-DHA)可以显着改善电泳DNA分离。 DNA电泳的单分子视频显微镜图像揭示了LPA-co-DHA基质中的新型链动力学,类似于蠕虫运动,我们将其归因于提高的DNA分辨率。尤其是在接近链间缠结阈值的聚合物/共聚物浓度下的LPA-co-DHA溶液中,DNA峰分离得到了显着改善。较高的聚合物浓度仅对较小的DNA分子(<120个碱基对)产生增强的分离。疏水性交联网络相对于基于增强分离性能(或速度)的传统线性聚合物和化学交联凝胶具有优势,因为疏水性交联可以可逆地断裂,从而使微通道负载更容易。

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