Sulfonated 4,7-dichlororhodamine dyes and their application

Abstract

The invention describes the preparation and use of sulfonated 4,7-dichlororhodamine dyes having the structure: Wherein R, R, Rand Rare independently hydrogen, —SOY where Y is H or a counterion, C-Calkyl, or C-Calkoxy, where each alkyl or alkoxy is optionally further substituted by a carboxylic acid, a salt of carboxylic acid, or a carboxylic acid ester of a C-Calcohol; Rand Rare independently hydrogen, C-Calkyl, C-Ccarboxyalkyl, C-Csulfoalkyl, a salt of C-Ccarboxyalkyl, or a salt of C-Csulfoalkyl; or Rin combination with R, or Rin combination with R, or both, form a 5- or 6-membered ring that is saturated or unsaturated, and is optionally substituted by one or more C-Calkyls, or —CHSOY moieties; Rand Rare independently hydrogen, C-Calkyl, C-Ccarboxyalkyl, C-Csulfoalkyl, a salt of C-Ccarboxyalkyl, or a salt of C-Csulfoalkyl; or Rin combination with R, or Rin combination with R, or both, form a 5- or 6-membered ring that is saturated or unsaturated, and is optionally substituted by one or more C-Calkyls, or —CHSOY moieties; Ris carboxylate or sulfonate; Rand Rare independently hydrogen, Cl, or R, where Ris a reactive group. In one aspect, the invention includes reagents labeled with the sulfonated 4,7-dichlororhodamine dye compounds, including an amino acid, peptide, protein, antibody, polysaccharide, nucleotide, nucleic acid, hapten, drug, lipid, polymer, cell, bacterium, yeast, or virus. In an additional aspect, the invention includes methods utilizing such dye compounds and reagents including protein labeling, dideoxy polynucleotide sequencing and fragment analysis methods.