聚合物溶液中DNA毛细管电泳的场效应和浓度效应的系统研究。

Systematic study of field and concentration effects in capillary electrophoresis of DNA in polymer solutions.

作者信息

Mitnik L, Salomé L, Viovy J L, Heller C

机构信息

Laboratoire de PhysicoChimie Théorique (URA CNRS 1382), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris, France.

出版信息

J Chromatogr A. 1995 Sep 1;710(2):309-21. doi: 10.1016/0021-9673(95)00500-5.

DOI:10.1016/0021-9673(95)00500-5

PMID:7550964

Abstract

A systematic study of the separation of double-stranded DNA in hydroxypropylcellulose (HPC) with a molecular mass of 10(6) was undertaken, using a variety of concentrations (from 0.1 to 1%) and different electric fields (from 6 to 540 V/cm). The data show that at high polymer concentrations ( > or = 0.4%) and low fields, the separation mechanism is similar to that occurring in gels. The results are in good agreement with theoretical models, and in particular with a recently proposed theory for gels with a pore size smaller than the persistence length of DNA. For more dilute solutions and high fields, however, the separation pattern cannot be explained by existing theories. The existence of an original mechanism was confirmed by the direct observation of the conformation of double-stranded DNA molecules in the polymer solution by fluorescence videomicroscopy. Practical conclusions for the capillary electrophoretic separation of duplex DNA are drawn.

摘要

对分子量为10⁶的羟丙基纤维素（HPC）中双链DNA的分离进行了系统研究，使用了多种浓度（从0.1%到1%）和不同电场（从6到540 V/cm）。数据表明，在高聚合物浓度（≥0.4%）和低电场下，分离机制与凝胶中的相似。结果与理论模型高度吻合，特别是与最近提出的孔径小于DNA持久长度的凝胶理论相符。然而，对于更稀的溶液和高电场，现有的理论无法解释分离模式。通过荧光视频显微镜直接观察聚合物溶液中双链DNA分子的构象，证实了一种新机制的存在。得出了双链DNA毛细管电泳分离的实际结论。