The characterization of composite agarose/hydroxyethylcellulose matrices for the separation of DNA fragments using capillary electrophoresis

Electrophoresis
B A SilesM F Warder

Abstract

Mixtures of the polysaccharide derivatives, 19% hydroxyethylated SeaPrep agarose (SP-AG) and hydroxyethylcellulose (HEC), in aqueous buffer solutions are applied for the first time to the separation of DNA fragments using capillary electrophoresis (CE). These matrices form unique size-sieving networks that allow the separation of a wide size range of DNA fragments in a single analysis. Relative to their homogeneous counterparts, the composite separation matrices provide enhanced selectivity properties of DNA fragments, especially for fragments greater than 1000 base pairs (bp) in length. Additionally, the effects on separation performance of capillary temperature, the incorporation of a DNA intercalator, and applied field strength are demonstrated. Solution viscosity measurements of the homogeneous and composite matrix solutions were made in order to establish the entanglement threshold concentrations for the unique size-sieving solutions. The relatively low solution viscosities of the composite separation matrices allow reproducible replacement of the separation matrix between analyses. The mechanism of separation of DNA fragments for the composite matrices is proposed.

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Citations

Feb 24, 2001·Electrophoresis·M N Albarghouthi, A E Barron
Jul 19, 2011·Analytical and Bioanalytical Chemistry·Zhenqing LiYoshinori Yamaguchi
Sep 7, 2002·Journal of Chromatography. a·Benjamin Chu, Dehai Liang
Jun 29, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·S A NevinsZ E Nackerdien
Jan 8, 2000·Journal of Biochemical and Biophysical Methods·P G Righetti, C Gelfi
Oct 12, 2010·Electrophoresis·Zhenqing LiYoshinori Yamaguchi
Feb 13, 2010·Electrophoresis·Kun-Hong LeeHuan-Tsung Chang
Jan 30, 2014·Journal of Chromatography. a·Zhenqing LiYoshinori Yamaguchi
Dec 3, 2014·Analytical Biochemistry·Zhenqing LiYoshinori Yamaguchi

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