Siles B A, Anderson D E, Buchanan N S, Warder M F
The College of William and Mary, Department of Chemistry, Williamsburg, VA 23187-8795, USA.
Electrophoresis. 1997 Oct;18(11):1980-9. doi: 10.1002/elps.1150181117.
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.
首次将多糖衍生物、19%羟乙基化的海普瑞琼脂糖（SP-AG）和羟乙基纤维素（HEC）的混合物应用于水性缓冲溶液中，通过毛细管电泳（CE）分离DNA片段。这些基质形成独特的尺寸筛分网络，能够在单次分析中分离出宽尺寸范围的DNA片段。相对于其均相对应物，复合分离基质提供了增强的DNA片段选择性，特别是对于长度大于1000碱基对（bp）的片段。此外，还展示了毛细管温度、DNA嵌入剂的加入以及施加场强对分离性能的影响。对均相和复合基质溶液进行了溶液粘度测量，以确定独特尺寸筛分溶液的缠结阈值浓度。复合分离基质相对较低的溶液粘度使得在分析之间能够可重复地更换分离基质。提出了复合基质分离DNA片段的机制。
