Fta. Chen et al., SEMICONDUCTOR LASER-INDUCED FLUORESCENCE DETECTION IN CAPILLARY ELECTROPHORESIS USING A CYANINE DYE, Journal of chromatography, 652(2), 1993, pp. 355-360
Cy5, an activated carboxyl cyanine fluorophore, was characterized by c
apillary 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 dicar
boxylic acid. A Cy5-labeled oligonucleotide M(13) primer for DNA seque
ncing (M13mp18 template) was synthesized with a purity of better than
95%. The labeled primer was analyzed by liquid chromatography, using U
V-visible detection, and by CE, monitored by laser-induced fluorescenc
e (LIE) detection. Analysis of the Cy5-labeled oligonucleotide primer
by CE-LIF in a 9% polyacrylamide gel-filled capillary indicated the pu
rity 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.