Transient orientation of linear DNA molecules during pulsed-field gel electrophoresis
Transient orientation of linear DNA molecules during pulsed-field gel electrophoresis
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脉冲场凝胶电泳期间线性 DNA 分子的瞬时取向
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Departnent
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作者：
G. Holzwarth;Chad B. McKee;Susan Steiger;Glenn Crater;Departnent
在脉冲场凝胶电泳过程中，测量了λ DNA和λ-DNA寡聚体的瞬时取向。DNA变得基本上平行于电场E排列。在响应单个矩形脉冲时，方向显示过冲，峰值在1秒，然后是小的下冲，最后是平稳状态。当磁场关闭时，取向以两个不同的指数阶段消散。场反转导致取向周期，当链反转方向时，具有减少取向的中间周期。场反转脉冲施加到线性寡聚体的DNA的取向响应减慢，但增加幅度的分子量增加，对于一个给定的字段。因为预期DNA拉伸和平行于E的排列与DNA速度相关，所以响应于脉冲场的速度也预期表现出过冲。
The transient orientation of lambda DNA and lambda-DNA oligomers has been measured during pulsed field gel electrophoresis. The DNA becomes substantially aligned parallel to the electric field E. In response to a single rectangular pulse, orientation shows an overshoot with a Peak at 1 second, then a small undershoot, and finally a plateau. When the field is turned off, the orientation dissipates in two distinct exponential phases. Field inversion leads to periods of orientation with intervening periods of reduced orientation as the chains reverse direction. Field inversion pulses applied to linear oligomers of lambda-DNA show that orientation responses slow down but increase in amplitude as molecular weight increases, for a given field. Because DNA stretching and alignment parallel to E are expected to correlate with DNA velocity, the velocity in response to a pulsed field is also expected to exhibit an overshoot.