Side-entry excitation and detection of square capillary array electrophoresis for DNA sequencing.
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.
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Ames Laboratory-US Department of Energy, Iowa State University, Ames, IA 50011, USA.
Yeung
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D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D008956:Models, Chemical; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*