Hightower R C, Metge D W, Santi D V
Department of Biochemistry, University of California, San Francisco 94143.
Nucleic Acids Res. 1987 Oct 26;15(20):8387-98. doi: 10.1093/nar/15.20.8387.
The migration properties of a series of supercoiled plasmids ranging in size from 4 to 16 kilobases (kb) have been analyzed by orthogonal-field-alternation gel electrophoresis (OFAGE). These circular DNAs enter the gel and are well resolved. Unlike linear DNA molecules, the relative mobilities of these plasmids are constant over a wide range of pulse times, from 10 to 120 seconds, as well as over a broad range of total running times, from 6 to 24 hours. Electrophoresis of supercoiled, relaxed, and nicked open circular forms as well as topoisomers of pBR322 shows that the extent of supercoiling has a dramatic effect on plasmid migration on OFAGE. Several practical applications for exploiting the different migration properties of circular and linear DNA molecules on OFAGE are presented.
通过正交交变电场凝胶电泳（OFAGE）分析了一系列大小从4到16千碱基（kb）的超螺旋质粒的迁移特性。这些环状DNA进入凝胶并得到很好的分离。与线性DNA分子不同，这些质粒的相对迁移率在很宽的脉冲时间范围（从10到120秒）以及总运行时间范围（从6到24小时）内都是恒定的。对pBR322的超螺旋、松弛和缺口开环形式以及拓扑异构体进行电泳表明，超螺旋程度对OFAGE上的质粒迁移有显著影响。本文还介绍了利用环状和线性DNA分子在OFAGE上不同迁移特性的几个实际应用。
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