title: Versatile procedure of multiple introduction of 8-aminomethylene blue into oligonucleotides.
abstract: The coupling of 8-aminomethylene blue to oligonucleotides via poly-L-glutamic acid linker using carboxy-anchor groups will be described. The introduction of carboxy-anchor groups into oligonucleotides proceeds both during automated synthesis using 6-(ethoxycarbonyl)hexyl 1-O-phosphoramidite and by reaction of 5'-amino-functionalized oligonucleotides with succinic anhydride. O-(N-Succinimidyl)-1,1,3,3-tetramethyluronium tetrafluoroborate was used as activating reagent for binding of poly-L-glutamic acid to the carboxylated oligonucleotides. The successful 5'-carboxylation and poly-L-glutamic acid coupling were proven both by polyacrylamide gel electrophoreses and HPLC. 8-Aminomethylene blue in its leucoform was covalently coupled to the oligonucleotides in the presence of water soluble carbodiimide.
authors.affiliation: Humboldt-Universität zu Berlin, Institut für Chemie, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Möller
authors.forename: F
authors.initials: F
authors.lastname: Schubert
authors.forename: D
authors.initials: D
authors.lastname: Cech
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008751:Methylene Blue / Q000737:chemistry*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D011099:Polyglutamic Acid / Q000737:chemistry; D013055:Spectrophotometry, Infrared
chemical_list: D007202:Indicators and Reagents; D009841:Oligonucleotides; D011099:Polyglutamic Acid; D008751:Methylene Blue