高效毛细管线性聚丙烯酰胺凝胶电泳柱的制备、柱性能表征及操作条件考察  

Preparation and Characterization of High Performance Linear Polyacrylamide Capillary Gel Electrophoresis Columns and Investigation of the Experimental Conditions

作　　者：施维[1] 张玉奎[1] 陈农[1] 张津[1] 王磊[1] 邹汉法[1] 

机构地区：[1]中国科学院大连化学物理研究所,国家色谱研究分析中心大连116011

出　　处：《分析化学》1998年第1期1-6,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金!(No.29575213);中国科学院基金资助项目

摘　　要：发展一种简便快速的线性聚丙烯酰胺毛细管凝胶电泳（ＣＧＥ）柱的新型制备方法，用于分离Ｐｏｌｙ ｄＡ（４０～６０）和双链ＤＮＡ，柱效达 ６百万理论板／米。提出“筛分能力”作为 ＣＧＥ柱评价指标，对样品迁移行为随操作条件的变化规律进行考察，为毛细管凝胶电泳的理论研究和实验条件优化奠定了基础。Linear polyacrylamide capillary gel electrophoresis (CGE) column is easier to prepare and can be used for a longer time, as compared with cross-linking gel column. A convenient and rapid method was developed to prepare isocratic and concentration gradient linear polyacrylamide columns, and poly dA(40-60) and double string DNA were separated efficiently with an efficiency of 6 × 106 plates/m. The characterization index of sieving power was proposed to evaluate the columns. The effects of experimental conditions including electric field strength and column temperature on the migration behavior were investigated. The peak width at half height was found to be linear with migration time under both Ogston and Reptation models. On concentration gradient gel column, the sample zone dispersion was smaller than that of isocratic gel column. These lay a foundation for the optimization and theoretical research of CGE both of isocratic and gradient modes.

关 键 词：毛细管凝胶电泳 柱制备 CGE柱 DNA 序列分析 

分 类 号：O657.8[理学—分析化学] Q523.03[理学—化学]