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Continuous, on‐line DNA sequencing using a versatile infrared laser scanner/electrophoresis apparatus
Author(s) -
Middendorf Lyle R.,
Bruce John C.,
Bruce Robert C.,
Eckles Robert D.,
Grone Daniel L.,
Roemer Stephen C.,
Sloniker Gary D.,
Steffens David L.,
Sutter Scott L.,
Brumbaugh John A.,
Patonay Gabor
Publication year - 1992
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501301103
Subject(s) - fluorophore , microscope , laser scanning , infrared , fluorescence , materials science , fluorescence microscope , laser , analytical chemistry (journal) , chemistry , optics , chromatography , physics
A new apparatus for continuously detecting fluorescently labeled DNA fragments is based on infrared fluorescence technology. This technology combines state‐of‐the‐art developments in chemistry, laser technology, and detection, while achieving improved reliability, sensitivity, and flexibility for applications including DNA sequencing. DNA molecules labeled with a novel infrared fluorophore are detected during electrophoresis using a scanning infrared fluorescence microscope. The microscope consists of a laser diode for exciting the fluorophore and a silicon avalanche photodiode for detecting the infrared emission. Optimum conditions for detection and throughput are obtained by adjusting electrophoresis, scanning and imaging parameters. Typical DNA sequencing runs (test templates) allow identification of over 500 bases per sample with > 99% accuracy.