Electromigration behavior of nucleic acids in capillary electrophoresis under pulsed-field conditions
Electromigration behavior of nucleic acids in capillary electrophoresis under pulsed-field conditions
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脉冲场条件下毛细管电泳中核酸的电迁移行为
DOI：
10.1016/j.chroma.2014.01.021
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发表时间：
2014-02-28
影响因子：
4.1
通讯作者：
Yamaguchi, Yoshinori
中科院分区：
文献类型：
--
作者：
Li, Zhenqing;Liu, Chenchen;Yamaguchi, Yoshinori
研究了脉冲电场条件下双链DNA(DsDNA)和RNA的迁移模式。通过计算核酸迁移率与其分子大小的双对数图，我们发现：(1)dsDNA分子遵循S型迁移模式，这可能与Ogston筛选、过渡模式和翻转模型有关。此外，如果DNA在较高分子质量的HEC中溶解，则过渡区域消失。(Ii)RNA的迁移模式与用于分离的变性剂有关。当RNA被醋酸变性时，其迁移率随其分子大小呈抛物线下降。以尿素为变性剂时，迁移率随分子大小呈线性下降。(3)尿素变性时，RNA可能以Ogston、Reptation方式迁移，没有取向机制；而乙酸变性时，这两种模型不适用于RNA。尽管PFCE的电泳条件不同，但仍能观察到S形、线性、抛物线的迁移模式。(4)在一定的调制深度下，醋酸的迁移时间(T-m)随平均分离电压(V-a)的增加而减小，当RNA在4.0M尿素中变性时，T-m与V-a呈线性相关。(V)由于聚合物的粘度降低，核酸在毛细管体积中的流动性随着人工温度的增加而增加。这是首次对脉冲电场作用下的大分子核酸进行系统的比较研究，为我们深入了解脉冲电场条件下RNA和DNA的迁移行为提供了新的思路。(C)2014爱思唯尔B.V.保留所有权利。
We have presented a study focused on the migration pattern of double-stranded DNA (dsDNA) and RNA under pulsed field conditions. By calculating the dependence of nucleic acid mobility on its molecular size in a double logarithm plot, we found that (I) dsDNA molecules proceeded by a sigmoidal migration regime which was probably related to Ogston sieving, transition regime, and reptation model. Furthermore, the transition regime disappeared if DNA was resolved in a higher molecular mass HEC. (II) The migration pattern of RNA was relevant to the denaturant used for separation. When RNA was denatured by acetic acid, its mobility parabolically declined with its molecular size. The mobility was linearly decreased with the molecular size if urea was employed as denaturant. (III) RNA may migrate by Ogston, reptation without orientation mechanism when denatured by urea, whereas these two models were not suitable for RNA if denatured by acetic acid. Even though the electrophoretic conditions of PFCE were varied, the sigmoidal, linear, parabolic migration patterns could still be observed. (IV) Under certain modulation depth, the migration time (T-m) of acetic acid decreased with the increase of average separation voltage (V-a), and when RNA denatured in 4.0 M urea, T-m showed a linear correlation with V-a. (V) The mobility of nucleic acids increased with the growth of artificial temperature in the capillary volume due to the decrease in the viscosity of the polymer. This is the first systematic and comparative research of high molecular mass nucleic acids in PFCE, which provides us deep insight into RNA and DNA migration behavior under pulsed electric field conditions. (C) 2014 Elsevier B.V. All rights reserved.