title: Practical synthesis of isomerically pure 5- and 6-carboxytetramethylrhodamines, useful dyes for DNA probes.
abstract: Simple and scalable synthesis of 5- and 6-carboxytetramethylrhodamines (TAMRAs) is reported. Acylation of 3-dimethylaminophenol with 1,2,4-benzenetricarboxylic anhydride afforded a mixture of 4-dimethylamino-2-hydroxy-2',4'(5')-dicarboxybenzophenones, which can be easily separated into individual compounds upon recrystallization from methanol and acetic acid. Individual benzophenones were reacted with 3-dimethylaminophenol to give 5- or 6-carboxytetramethylrhodamines. The dyes were converted into hydroxyprolinol-based phosphoramidite reagents suitable for oligonucleotide synthesis. 5- and 6-TAMRA isomers on oligonucleotides showed similar absorption and emission spectra. Fluorescence quantum yield of the dyes correlates with the presence of dG nucleosides in the adjacent region of oligonucleotide sequence. Several energy transfer primers containing on their 5'-termini (6-FAM)dT(n)(6-TAMRA) dye system (n = 0, 2, 4, 6, 8, 10, 12, 14) were prepared, and their spectral properties were studied.
authors.affiliation: Institute of Physical Organic Chemistry, Minsk, Belarus.
authors.forename: Maksim V
authors.initials: MV
authors.lastname: Kvach
authors.forename: Irina A
authors.initials: IA
authors.lastname: Stepanova
authors.forename: Igor A
authors.initials: IA
authors.lastname: Prokhorenko
authors.forename: Aleksander P
authors.initials: AP
authors.lastname: Stupak
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Bolibrukh
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Korshun
authors.forename: Vadim V
authors.initials: VV
authors.lastname: Shmanai
mesh_terms: D015342:DNA Probes / Q000138:chemical synthesis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D007536:Isomerism; D015394:Molecular Structure; D012235:Rhodamines / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D012235:Rhodamines; C005358:tetramethylrhodamine