通过毛细管电泳实现线性和超螺旋DNA的高速分离。缓冲液、缠结聚合物及电场效应。

High-speed separation of linear and supercoiled DNA by capillary electrophoresis. Buffer, entangling polymer, and electric field effects.

作者信息

Oana H, Hammond R W, Schwinefus J J, Wang S C, Doi M, Morris M D

机构信息

Department of Chemistry, University of Michigan, Ann Arbor 48109, USA.

出版信息

Anal Chem. 1998 Feb 1;70(3):574-9. doi: 10.1021/ac970938j.

DOI:10.1021/ac970938j

PMID:9470487

Abstract

Capillary electrophoresis in dilute and semidilute (slightly entangled) hydroxyethyl cellulose (HEC) is shown to separate linear double-stranded DNA (ds-DNA) and supercoiled plasmid DNA in the size range 1-16 thousand base pairs in 3 min. The mobilities of linear ds-DNA fragments are stronger functions of electric field strength and buffer concentration than the mobilities of supercoiled plasmids. The effects of HEC concentration and molecular weight are similar for both forms of DNA. The behavioral differences, which are attributed to the greater stiffness of the plasmids, can be used to define conditions that maximize resolution of supercoiled and linear ds-DNA of the same or similar number of base pairs.

摘要

在稀释和半稀释（轻度缠结）的羟乙基纤维素（HEC）中进行毛细管电泳，结果表明在3分钟内可分离出大小范围为1至16千碱基对的线性双链DNA（ds-DNA）和超螺旋质粒DNA。线性ds-DNA片段的迁移率比超螺旋质粒的迁移率受电场强度和缓冲液浓度的影响更大。HEC浓度和分子量对两种形式的DNA的影响相似。这些行为差异归因于质粒更大的刚性，可用于确定使相同或相似碱基对数的超螺旋和线性ds-DNA分辨率最大化的条件。