毛细管电泳-激光诱导荧光鞘流检测系统优化研究及在基因分析中的应用  

Optimization of Capillary Electrophoresis-Laser Induced Fluorescence Detection with Sheath-Flow Cuvette and Its Application to Genetic Analysis

作　　者：陈林[1] 任吉存[1] 

机构地区：[1]上海交通大学化学化工学院,上海200240

出　　处：《分析化学》2003年第12期1413-1416,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金 (No .2 0 2 710 3 3 );国家自然科学基金重点项目 (No .2 0 3 3 5 0 2 0 );科技部 863重大专项 (No .2 0 0 2AA43 13 10 );上海市自然科学基金 (No .0 2ZA14 0 5 7);教育部和人事部留学归国人员启动基金资助

摘　　要：对自组装的毛细管电泳 激光诱导荧光鞘流检测装置进行了系统的优化。探讨了鞘流速度、激光功率和检测的位置对检测信号的影响以及鞘流速度与峰面积和理论塔板数的关系。在优化的条件下 ,对荧光素检测的线性范围为 1× 1 0 - 8～1× 1 0 - 1 0 mol L;检出限为 7.5×1 0 - 1 1 mol L(S N =3 )。使用聚N ,N 二甲基丙烯酰胺 (PDMA)为双功能非胶筛分介质 ,将毛细管电泳 激光诱导荧光鞘流检测系统成功地用于DNA片段及基因PCR扩增产物分离检测。We systematically investigated the effects of some factors, such as the sheath-flow velocity, laser power and detection position, on capillary electrophoresis-laser induced fluorescence detection (CE-LIF) with a sheath-flow cuvette. The data demonstrated that the detection signals were markedly decreasing, but the numbers of theoretical plate were significantly increasing with an increase in the sheath-flow. In the low laser power, the detection signals linearly increased with laser power. Under the optimum conditions, CE-LIF has a wide linear range from 1x10(-8) to 1x10(-10) mol/L, and a low detection limit of 7.5x10(-11) mol/L (S/N=3) for fluorescein. This system was. successfully used for the detection of deoxyribonucleic acid (DNA) fragments and polymerase chain reaction (PCR) products using polydimethylacrylamide as a sieving medium.

关 键 词：毛细管电泳-激光诱导荧光鞘流检测系统 优化 基因分析 鞘流速度 激光功率 峰面积 理论塔板数 

分 类 号：O658.9[理学—分析化学] Q523[理学—化学]