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凝胶电泳中的谱带展宽:通过荧光恢复后光漂白(FRAP)测量的扩散系数的标度律

Band broadening in gel electrophoresis: scaling laws for the dispersion coefficient measured by FRAP.

作者信息

Tinland B, Pernodet N, Pluen A

机构信息

Institut Charles Sadron, CNRS, Strasbourg, France.

出版信息

Biopolymers. 1998 Oct 5;46(4):201-14. doi: 10.1002/(SICI)1097-0282(19981005)46:4<201::AID-BIP2>3.0.CO;2-T.

Abstract

We determined quantitatively the band broadening effect during gel electrophoresis by measuring the longitudinal dispersion coefficient Dx, with a fluorescence recovery after photobleaching setup, coupled to an electrophoretic cell. We carried out measurements as a function of the electric field, the average pore size, and the molecular length of DNA fragments. Our results are in good agreement with the predictions of the biased reptation model with fluctuations described by T. A. Duke et al. [(1992) Physics Review Letters, vol. 69, pp. 3260-3263]. This agreement is observed on single-stranded DNA [persistence length approximately equal to 4 nm; B. Tinland et al. (1997) Macromolecules, vol. 30, pp. 5763-5765] in polyacrylamide gels and on double-stranded DNA (persistence length approximately equal to 50 nm) in agarose gels, two systems where the ratio between the average pore size and the Kuhn length is larger than 1.

摘要

我们通过测量纵向扩散系数Dx,利用光漂白后荧光恢复装置与电泳槽相结合的方法,对凝胶电泳过程中的谱带展宽效应进行了定量测定。我们根据电场、平均孔径和DNA片段的分子长度进行了测量。我们的结果与T. A. Duke等人[(1992年)《物理评论快报》,第69卷,第3260 - 3263页]所描述的具有涨落的偏置爬行模型的预测结果高度吻合。在聚丙烯酰胺凝胶中的单链DNA[持久长度约为4纳米;B. Tinland等人(1997年)《大分子》,第30卷,第5763 - 5765页]以及琼脂糖凝胶中的双链DNA(持久长度约为50纳米)上均观察到了这种吻合,这两个系统中平均孔径与库恩长度的比值大于1。

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