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超稀聚合物溶液中毛细管电泳分离DNA的瞬态纠缠耦合机制

A transient entanglement coupling mechanism for DNA separation by capillary electrophoresis in ultradilute polymer solutions.

作者信息

Barron A E, Blanch H W, Soane D S

机构信息

Department of Chemical Engineering, University of California, Berkeley 94720.

出版信息

Electrophoresis. 1994 May;15(5):597-615. doi: 10.1002/elps.1150150184.

Abstract

Using capillary electrophoresis, large DNA molecules (2.0-23.1 kbp) may be rapidly separated in ultradilute polymer solutions (< 0.002% w/w) under a high-voltage, steady field (265 V/cm). At this polymer concentration, the separation mechanism appears to be significantly different from that postulated to occur in crosslinked gels. Based on experimental results obtained with DNA restriction fragments and with negatively charged latex microspheres, we conclude that the Ogston and reptation models typically used to describe gel electrophoresis are not appropriate for DNA separations in such dilute polymer solutions. Electrophoresis experiments employing solutions of both small and large hydroxyethyl cellulose polymers highlight the importance of polymer length and concentration for the optimum resolution of DNA fragments varying in size from 72 bp to 23.1 kbp. A transient entanglement coupling mechanism for DNA separation in dilute polymer solutions is developed, which suggests that there is no a priori upper size limit to DNA that can be separated by capillary electrophoresis in a constant field.

摘要

使用毛细管电泳,大的DNA分子(2.0 - 23.1千碱基对)可在超稀聚合物溶液(<0.002% w/w)中,于高电压、稳定电场(265伏/厘米)下快速分离。在此聚合物浓度下,分离机制似乎与交联凝胶中假定发生的机制显著不同。基于用DNA限制片段和带负电荷的乳胶微球获得的实验结果,我们得出结论,通常用于描述凝胶电泳的奥格斯顿模型和蛇行模型不适用于此类稀聚合物溶液中的DNA分离。使用大小不同的羟乙基纤维素聚合物溶液进行的电泳实验突出了聚合物长度和浓度对于最佳分离大小从72碱基对到23.1千碱基对不等的DNA片段的重要性。我们提出了一种稀聚合物溶液中DNA分离的瞬态缠结耦合机制,这表明在恒定电场中通过毛细管电泳分离的DNA不存在先验的大小上限。

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