Lu S X, Yeung E S
Ames Laboratory-US Department of Energy, Iowa State University, Ames, IA 50011, USA.
J Chromatogr A. 1999 Aug 20;853(1-2):359-69. doi: 10.1016/s0021-9673(99)00676-7.
In high throughput DNA sequencing based on capillary electrophoresis, efficient coupling of the laser to each capillary is a challenge. Our group previously reported two multiple point irradiation schemes. The present work describes a more efficient excitation and detection method in which the laser light propagates through the capillary array without undergoing a serious reduction in power. An array of square capillaries (340 microns O.D. x 75 microns I.D.) was sandwiched between two fused-silica plates with an index-matching solution in between. The light was directed into the channel across the capillary array from the side. DNA sequences of PGEM/U from 24 capillaries were obtained even with a relatively low-power laser. The excitation scheme can be scaled up to hundreds of capillaries to achieve high-speed, high-throughput DNA sequencing, genetic typing and drug screening.
在基于毛细管电泳的高通量DNA测序中，实现激光与每根毛细管的高效耦合是一项挑战。我们小组之前报道了两种多点照射方案。目前的工作描述了一种更高效的激发和检测方法，其中激光在毛细管阵列中传播时功率不会严重降低。将一排方形毛细管（外径340微米×内径75微米）夹在两块熔融石英板之间，中间填充折射率匹配溶液。光从侧面射向穿过毛细管阵列的通道。即使使用相对低功率的激光，也能从24根毛细管中获得PGEM/U的DNA序列。该激发方案可扩展到数百根毛细管，以实现高速、高通量的DNA测序、基因分型和药物筛选。
