Nucleic acids, separation electrophoresis

Analysis of DNA or RNA frequently starts with the size separation of extracted nucleic acids by electrophoresis on agarose or... 

Nucleic acids can be visualized on the slab gel after separation by soaking in a solution of ethidium bromide, a dye that displays enhanced fluorescence when intercalated between stacked nucleic acid bases. Ethidium bromide may be added directly to the agarose solution before gel formation. This method allows monitoring of nucleic acids during electrophoresis. Irradiation of ethidium bromide-treated gels by UV light results in orange-red bands where nucleic acids are present. 

The separation of nucleic acids by CE has become a steadily growing area of interest, especially since the inception of the Human Genome Initiative. This interest originally stemmed from the use of polyacrylamide (PA) or agarose slab gel electrophoresis as the accepted standard for nucleic acid separation (Stellwagen, 1987). 

How are nucleic acids separated Two of the primary necessities for successful experiments with nucleic adds are to separate the components of a mixture and to detect the presence of nucleic acids. DNA can be cut into pieces with restriction enzymes and then separated with gel electrophoresis. 

Noolandi, J. A new concept for separating nucleic-acids by electrophoresis in solution using hybrid synthetic end labeled nucleic-acid molecules. Electrophoresis 14, 680-681 (1993). 

Zabzdyr, J.L. and LiUard, S.J., UV- and visible-excited fluorescence of nucleic acids separated by capillary electrophoresis, J. Chromatogr. A, 911, 269, 2001. 

This review gives an overview of current advances in nucleic acid separation by CE and microchip electrophoresis. The focus is on the separation mechanisms during CE, conventional separation matrices and thermoresponsive polymers solutions, UV and fluorescence detection, microchip-based CE, and entropic trapping networks. 

Y. Baba, M. Tsuhako, Gel-filled capillaries for nucleic acid separations in capillary electrophoresis. Trends Anal. Chem., 1992, n, 280-287. 

Liu, M. Chen, F. Nucleic acid separation and detection by electrophoresis with a counter-migrating high-affinity intercalating dye. U.S. Pat. Appl. Publ. 

Classical gel electrophoresis has been used extensively for protein and nucleic acid purification and characterization [9, 10], but has not been used routinely for small molecule separations, other than for polypeptides. A comparison between TLC and electrophoresis reveals that while detection is usually accomplished off-line in both electrophoretic and TLC methods, the analyte remains localized in the TLC bed and the mobile phase is immediately removed subsequent to chromatographic development. In contrast, in gel electrophoresis, the gel matrix serves primarily as an anti-... 

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