SEPARATION OF BRANCHED FROM LINEAR DNA BY TWO-DIMENSIONAL GEL-ELECTROPHORESIS
SEPARATION OF BRANCHED FROM LINEAR DNA BY TWO-DIMENSIONAL GEL-ELECTROPHORESIS
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DOI：
10.1016/0003-2697(83)90628-0
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发表时间：
1983-01-01
影响因子：
2.9
通讯作者：
BYERS, B
中科院分区：
文献类型：
--
作者：
BELL, L;BYERS, B
发展了一种从线性形式中分离分支DNA分子（如复制叉和重组中间体）的通用方法。使用作为体外构建的稳定X形分子的模型[来自酿酒酵母DNA]，这种分支形式在琼脂糖凝胶电泳过程中的迁移速度比相同质量的线性形式更慢。更高的琼脂糖浓度和更高的电泳电压增强的延迟的程度。这些性质为通过改变琼脂糖浓度和电压在2维上分离分支分子和线性分子的电泳方法提供了基础。在第一维中，浓度和电压都很低;在第二维中，这两个参数都增加了，从而迫使X形分子迁移到与线性分子的对角线弧不同的位置。此外，二维电泳能够将不同质量的X形形式彼此分离，以及与线性分子分离。
A general method for separating branched DNA molecules, such as replication forks and recombination intermediates, from linear forms was developed. Using as a model of stable X-shaped molecule constructed in vitro [from Saccharomyces cerevisiae DNA], this branched form migrated more slowly during agarose gel electrophoresis than did a linear form of the same mass. Higher agarose concentrations and higher electrophoretic voltages enhanced the extent of retardation. These properties provided the basis for an electrophoretic method of separating branched from linear molecules by variation of agarose concentration and voltage over 2 dimensions. In the first dimension, concentration and voltage were low; in the second, both parameters were increased, thereby forcing X-shaped molecules to migrate to positions distinct from a diagonal arc of linear molecules. In addition, 2-dimensional electrophoresis was capable of separating X-shaped forms of different mass from each other, as well as from linear molecules.