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DNA电泳迁移率的场强和孔径依赖性:光漂白后荧光恢复与电双折射测量的结合

Field and pore size dependence of the electrophoretic mobility of DNA: a combination of fluorescence recovery after photobleaching and electric birefringence measurements.

作者信息

Tinland B, Pernodet N, Weill G

机构信息

Institut Charles Sadron, CNRS, University Louis Pasteur, Strasbourg, France.

出版信息

Electrophoresis. 1996 Jun;17(6):1046-51. doi: 10.1002/elps.1150170612.

Abstract

By combining an electrophoretic cell with a setup of fluorescence recovery after photobleaching (FRAP) we can measure the electrophoretic mobility mu of double-stranded lambda DNA in agarose gel as a function of electric field E and gel concentration C. Mobility varies linearly with the field in agreement with the biased reptation model with fluctuations. The slopes are analyzed in term of orientation and compared with birefringence results. The mobility extrapolated at zero field follows the prediction of the reptation theory; we deduced the variation of the pore size with the agarose concentration. With a special use of our setup, we measure directly the free-mobility mu 0 of the DNA.

摘要

通过将电泳槽与光漂白后荧光恢复(FRAP)装置相结合,我们可以测量琼脂糖凝胶中双链λDNA的电泳迁移率μ随电场E和凝胶浓度C的变化。迁移率与电场呈线性变化,这与带有涨落的偏置爬行模型一致。从取向角度分析斜率,并与双折射结果进行比较。在零场下外推得到的迁移率符合爬行理论的预测;我们推导了孔径随琼脂糖浓度的变化。通过对我们装置的特殊应用,我们直接测量了DNA的自由迁移率μ0。

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