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脉冲场凝胶电泳过程中线性DNA分子的瞬时取向

Transient orientation of linear DNA molecules during pulsed-field gel electrophoresis.

作者信息

Holzwarth G, McKee C B, Steiger S, Crater G

机构信息

Department of Physics, Wake Forest University, Winston-Salem, NC 27109.

出版信息

Nucleic Acids Res. 1987 Dec 10;15(23):10031-44. doi: 10.1093/nar/15.23.10031.

Abstract

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.

摘要

在脉冲场凝胶电泳过程中,已对λ噬菌体DNA和λ噬菌体DNA寡聚物的瞬时取向进行了测量。DNA基本平行于电场E排列。响应单个矩形脉冲时,取向会出现一个峰值在1秒的过冲,然后是一个小的下冲,最后达到平稳状态。当电场关闭时,取向以两个不同的指数阶段消散。场反转会导致取向周期,随着链反向,其间会有取向降低的时期。施加到λ噬菌体DNA线性寡聚物上的场反转脉冲表明,对于给定的场,随着分子量增加,取向响应会变慢,但幅度会增加。由于预计DNA的拉伸和与E平行的排列与DNA速度相关,因此预计响应脉冲场的速度也会出现过冲。

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