Fiber bundle based scanning detection system for automated DNA sequencing.
High-throughput DNA sequencing techniques are under rapid development currently, mainly triggered by the Human Genome Project. At the present time, slab gel based automated DNA sequencing is the standard procedure, utilizing fluorophore labeling and laser-induced fluorescence detection with scanning technology. In this paper, a novel, fiber-optic bundle based detection system is introduced, where a central illuminating fiber is used for the excitation of the electrophoretically separated fluorophore-labeled DNA sequencing fragments, along with several collecting fibers disposed around the illuminating fiber to collect the emitted fluorescent signal. As a model system, Cy5-labeled DNA sequencing fragments were separated on an ultrathin polyacrylamide slab gel and detected by the fiber bundle based laser-induced fluorescence detection system. A 640-nm diode laser was used to generate the illumination beam, and the emitted light collected by the fiber bundle was detected by a solid-state avalanche photodiode.
Trost
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P
Genetic BioSytems, Inc., San Diego, California 92121, USA.
Guttman
A
A
D001322:Autoanalysis; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005336:Fiber Optic Technology; D005470:Fluorometry / Q000295:instrumentation; D006801:Humans; D008969:Molecular Sequence Data; D055100:Optical Fibers; D017422:Sequence Analysis, DNA / Q000379:methods*