A set of 4,7-dichlororhodamine compounds useful as fluorescent dyes are
disclosed having the structures
##STR00001##
wherein R.sub.1-R.sub.6 are hydrogen, fluorine, chlorine, lower alkyl,
lower alkene, lower alkyne, sulfonate, sulfone, amino, amido, nitrile,
lower alkoxy, linking group, or, when taken together, R.sub.1 and R.sub.6
is benzo, or, when taken together, R.sub.4 and R.sub.5 is benzo;
R.sub.7-R.sub.10, R.sub.12-R.sub.16 and R.sub.18 may be hydrogen,
fluorine, chlorine, lower alkyl lower alkene, lower alkyne, sulfonate,
sulfone, amino, amido, nitrile, lower alkoxy, linking group; R.sub.11 and
R.sub.17 may be hydrogen, lower alkyl lower alkene, lower alkyne, phenyl,
aryl, linking group; Y.sub.1-Y.sub.4 are hydrogen, lower alkyl or
cycloalkyl, or, when taken together, Y.sub.1 and R.sub.2, Y.sub.2 and
R.sub.1 Y.sub.3 and R.sub.3, and/or Y.sub.4 and R.sub.4 is propano,
ethano, or substituted forms thereof, and X.sub.1-X.sub.3 taken
separately are hydrogen, chlorine, fluorine, lower alkyl, carboxylate,
sulfonate, hydroxymethyl, and linking group, or any combinations thereof.
In another aspect, the invention includes reagents labeled with the
4,7-dichlororhodamine dye compounds, including deoxynucleotides,
dideoxynucleotides, and polynucleotides. In an additional aspect, the
invention includes methods utilizing such dye compounds and reagents
including dideoxy polynucleotide sequencing and fragment analysis
methods.