Shi Y, Simpson P C, Scherer J R, Wexler D, Skibola C, Smith M T, Mathies R A
Chemistry Department, University of California, Berkeley 94720, USA.
Anal Chem. 1999 Dec 1;71(23):5354-61. doi: 10.1021/ac990518p.
The design, fabrication, and operation of a radial capillary array electrophoresis microplate and scanner for high-throughput DNA analysis is presented. The microplate consists of a central common anode reservoir coupled to 96 separate microfabricated separation channels connected to sample injectors on the perimeter of the 10-cm-diameter wafer. Detection is accomplished by a laser-excited rotary confocal scanner with four color detection channels. Loading of 96 samples in parallel is achieved using a pressurized capillary array system. High-quality separations of 96 pBR322 restriction digest samples are achieved in < 120 s with the microplate system. The practical utility and multicolor detection capability is demonstrated by analyzing 96 methylenetetrahydrofolate reductase (MTHFR) alleles in parallel using a noncovalent 2-color staining method. This work establishes the feasibility of performing high-throughput genotyping separations with capillary array electrophoresis microplates.
介绍了一种用于高通量DNA分析的径向毛细管阵列电泳微孔板及扫描仪的设计、制造和操作。该微孔板由一个中央公共阳极储液器组成，该储液器与96个独立的微制造分离通道相连，这些通道连接到直径为10厘米的晶圆周边的样品注入器上。检测通过具有四个颜色检测通道的激光激发旋转共聚焦扫描仪完成。使用加压毛细管阵列系统可实现96个样品的并行加载。该微孔板系统在不到120秒的时间内实现了96个pBR322限制性消化样品的高质量分离。通过使用非共价双色染色方法并行分析96个亚甲基四氢叶酸还原酶（MTHFR）等位基因，证明了其实际效用和多色检测能力。这项工作确立了用毛细管阵列电泳微孔板进行高通量基因分型分离的可行性。
Zotero 插件