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. 1993 Oct 22;652(2):355-60.
doi: 10.1016/0021-9673(93)83253-o.

Semiconductor laser-induced fluorescence detection in capillary electrophoresis using a cyanine dye

F T Chen  1 , A Tusak, S Pentoney Jr, K Konrad, C Lew, E Koh, J Sternberg
Affiliations

Semiconductor laser-induced fluorescence detection in capillary electrophoresis using a cyanine dye

F T Chen et al. J Chromatogr A. .

Abstract

Cy5, an activated carboxyl cyanine fluorophore, was characterized by capillary electrophoresis (CE) using a semiconductor laser at 652 nm to induce fluorescence. Hydrolysis of the activated Cy5 in the presence of ammonia results in the formation of a mono- and diamide and a dicarboxylic acid. A Cy5-labeled oligonucleotide M13 primer for DNA sequencing (M13mp18 template) was synthesized with a purity of better than 95%. The labeled primer was analyzed by liquid chromatography, using UV-visible detection, and by CE, monitored by laser-induced fluorescence (LIF) detection. Analysis of the Cy5-labeled oligonucleotide primer by CE-LIF in a 9% polyacrylamide gel-filled capillary indicated the purity of the major Cy5-oligonucleotide primer was greater than 90%. The detection sensitivity for Cy5-based CE-LIF detection system with a 2.5-mW red semiconductor laser is about 10(-10) M.

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