SEPARATIONS OF OPEN-CIRCULAR DNA USING PULSED-FIELD ELECTROPHORESIS
SEPARATIONS OF OPEN-CIRCULAR DNA USING PULSED-FIELD ELECTROPHORESIS
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DOI：
10.1073/pnas.84.12.4054
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发表时间：
1987-06-01
影响因子：
11.1
通讯作者：
ZIMM, BH
中科院分区：
文献类型：
--
作者：
LEVENE, SD;ZIMM, BH
高电场对琼脂糖中开环 DNA 凝胶电泳迁移率的影响与对相同分子量的线性分子的影响显着不同。在高场下，足够大的环状 DNA 会被阻止迁移到凝胶中，而线性分子和较小的环状 DNA 会正常迁移。这种效应强烈依赖于场，随着场强的增加，影响尺寸减小的圆形分子。我们使用连续和反向脉冲电场，研究了一系列 2.9 至 56 kilobase 对的质粒 DNA 的这种效应。反向脉冲的应用消除了某些条件下的效应，并支持开环 DNA 的凝胶电泳模型，其中通过接合琼脂糖凝胶纤维的自由端来捕获环状形式。
The effect of high electric fields on the gel-electrophoretic mobility of open-circular DNA in agarose differs dramatically from that on linear molecules of the same molecular weight. At high fields, sufficiently large circular forms are prevented from migrating into the gel whereas linear molecules and smaller circular DNAs migrate normally. This effect is strongly field dependent, affecting circular molecules of decreasing size with increasing field strength. We have studied this effect with a series of plasmid DNAs ranging from 2.9 to 56 kilobase pairs using continuous and reversing-pulse electric fields. Application of reversing pulses abolishes the effect under certain conditions and supports the model for the gel electrophoresis of open-circular DNA where circular forms are trapped by engaging the free end of an agarose gel fiber.