Smith E M, Xu H, Ewing A G
Department of Chemistry, The Pennsylvania State University, University Park 16802, USA.
Electrophoresis. 2001 Jan;22(2):363-70. doi: 10.1002/1522-2683(200101)22:2<363::AID-ELPS363>3.0.CO;2-G.
A novel method is presented for automated injection of DNA samples into microfabricated separation devices via capillary electrophoresis. A single capillary is used to electrokinetically inject discrete plugs of DNA into an array of separation lanes on a glass chip. A computer-controlled micromanipulator is used to automate this injection process and to repeat injections into five parallel lanes several times over the course of the experiment. After separation, labeled DNA samples are detected by laser-induced fluorescence. Five serial separations of 6-carboxyfluorescein (FAM)-labeled oligonucleotides in five parallel lanes are shown, resulting in the analysis of 25 samples in 25 min. It is estimated that approximately 550 separations of these same oligonucleotides could be performed in one hour by increasing the number of lanes to 37 and optimizing the rate of the manipulator movement. Capillary sample introduction into chips allows parallel separations to be continuously performed in serial, yielding high throughput and minimal need for operator intervention.
提出了一种通过毛细管电泳将DNA样品自动注入微制造分离装置的新方法。使用单个毛细管通过电动方式将离散的DNA塞注入玻璃芯片上的一系列分离通道中。使用计算机控制的微操纵器使该注入过程自动化，并在实验过程中多次重复注入五个平行通道。分离后，通过激光诱导荧光检测标记的DNA样品。展示了在五个平行通道中对6-羧基荧光素（FAM）标记的寡核苷酸进行的五次连续分离，从而在25分钟内分析了25个样品。据估计，通过将通道数量增加到37并优化操纵器的移动速度，一小时内可以对这些相同的寡核苷酸进行大约550次分离。将毛细管样品引入芯片允许连续地串行进行平行分离，从而实现高通量且对操作员干预的需求最小。
