用于DNA脉冲场凝胶电泳分离的优化条件。

Birren B W, Lai E, Clark S M, Hood L, Simon M I

Division of Biology, California Institute of Technology, Pasadena 91125.

Nucleic Acids Res. 1988 Aug 11;16(15):7563-82. doi: 10.1093/nar/16.15.7563.

Quantitative measurement of DNA migration in gel electrophoresis requires precisely controlled homogeneous electric fields. A new electrophoresis system has allowed us to explore several parameters governing DNA migration during homogeneous field pulsed field gel (PFG) electrophoresis. Migration was measured at different switch times, temperatures, agarose concentrations, and voltage gradients. Conditions which increase DNA velocities permit separation over a wider size range, but reduce resolution. We have also varied the angle between the alternating electric fields. Reorientation angles between 105 degrees and 165 degrees give equivalent resolution, despite significant differences in DNA velocity. Separation of DNA fragments from 50 to greater than 7000 kilobases (Kb) can easily be optimized for speed and resolution based on conditions we describe.

凝胶电泳中DNA迁移的定量测量需要精确控制的均匀电场。一种新的电泳系统使我们能够探索在均匀场脉冲场凝胶（PFG）电泳过程中控制DNA迁移的几个参数。在不同的切换时间、温度、琼脂糖浓度和电压梯度下测量迁移情况。提高DNA速度的条件允许在更宽的大小范围内进行分离，但会降低分辨率。我们还改变了交变电场之间的角度。尽管DNA速度存在显著差异，但105度至165度之间的重新定向角度能提供等效的分辨率。根据我们描述的条件，可以很容易地针对速度和分辨率对50至大于7000千碱基（Kb）的DNA片段分离进行优化。