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.