Pulsed-field gel electrophoresis of circular DNA.
Pulsed-field gel electrophoresis of circular DNA.
复制标题
环状 DNA 的脉冲场凝胶电泳。
DOI：
10.1093/nar/17.11.4359
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发表时间：
1989
影响因子：
14.9
通讯作者：
Scherer,S
中科院分区：
文献类型：
--
作者：
Simske,JS;Scherer,S
超螺旋（I型）和切口环状（II型）质粒DNA的迁移率在两种主要形式的脉冲场电泳CHEF和OFAGE上测定。以Saccharomyces酵母染色体为标准品，用分子长度为2.30 ~ 17.8个酶对的质粒进行酶解。琼脂糖凝胶浓度从0.3%到2.0%不等，较高百分比的凝胶分辨较小质粒的I型和II型。3.7至240秒的脉冲范围导致在0.5%和1.0%凝胶上的酵母染色体迁移率相当可变，而所有质粒DNA的I型和II型都显示出相对恒定的迁移率，在最短的脉冲时间内有所增加。使用30秒的脉冲时间和至少1.0%的凝胶浓度，可以改变17.8kb质粒的质粒形式的通常迁移顺序。我们将此结果解释为在我们的脉冲场凝胶条件下II型的相对迁移率增加。
Mobility of supercoiled (form I) and nicked circular (form II) plasmid DNAs was determined on two major forms of pulsed-field electrophoresis, CHEF and OFAGE. Plasmids with molecular lengths ranging from 2.30 to 17.8 kilobase pairs (kb) were used withSac-charomyces cerevisiaechromosomes as standards. Agarose gel concentrations were varied from 0.3 to 2.0 percent, with higher percentage gels resolving forms I and II of smaller plasmids. The pulsing range of 3.7 to 240 seconds resulted in quite variable Saccharomyces chromosomal mobilities on both 0.5 and 1.0 percent gels, while both form I and II of all plasmid DNAs showed relatively constant mobilities with some increase at the shortest pulse times. Using a 30 second pulse time and gel concentrations of at least 1.0 percent, the usual order of migration of plasmid forms for a 17.8kb plasmid could be changed. We interpret this result as an increase in the relative mobility of form II in our pulsed-field gel conditions.