使用低凝胶、可替换琼脂糖凝胶对0.1 - 23 kbp DNA片段进行高性能场反转毛细管电泳。
High-performance field inversion capillary electrophoresis of 0.1-23 kbp DNA fragments with low-gelling, replaceable agarose gels.
作者信息
Chen N, Wu L, Palm A, Srichaiyo T, Hjertén S
机构信息
Department of Biochemistry, Uppsala University, Biomedical Center, Sweden.
出版信息
Electrophoresis. 1996 Sep;17(9):1443-50. doi: 10.1002/elps.1150170908.
Field inversion capillary gel electrophoresis (FICGE) has been used for the separation of 0.1-23 kbp DNA fragments in a low-melting, low-gelling agarose gel. The influence of the amplitude of the voltage pulses, the pulse times and gel concentrations on the separation factor has been studied and found to be similar to that of polyacrylamide gels. Very high resolution can be obtained by altering continuously the pulse times and/or voltage according to a program tailor-made for the size of the DNA molecules to be separated (programmed FICGE). The advantage of these agarose gels in comparison with polyacrylamide gels is that they are nontoxic, easy to prepare and they have high UV transmission. They give a resolution that is equivalent to or better than that of polyacrylamide gels, and the risk of bubble formation in the gel is small. In addition, they are replaceable and, therefore, very convenient for automated analyses.
场反转毛细管凝胶电泳（FICGE）已用于在低熔点、低凝胶化琼脂糖凝胶中分离0.1 - 23 kbp的DNA片段。研究了电压脉冲幅度、脉冲时间和凝胶浓度对分离因子的影响，发现其与聚丙烯酰胺凝胶的影响相似。通过根据为待分离DNA分子大小量身定制的程序（程序化FICGE）连续改变脉冲时间和/或电压，可以获得非常高的分辨率。与聚丙烯酰胺凝胶相比，这些琼脂糖凝胶的优点是无毒、易于制备且具有高紫外透射率。它们给出的分辨率与聚丙烯酰胺凝胶相当或更好，并且凝胶中形成气泡的风险较小。此外，它们可更换，因此非常便于自动化分析。
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