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• Cyanine3 phosphoramidite 5'

Cyanine3 phosphoramidite 5'

Cyanine3 is a fluorophore that is widely used in molecular biology experiments such as oligonucleotide labeling followed by oligonucleotide detection. By its spectral characteristics, Cyanine3 is an analog of Cy™3 with a fluorescence maximum at 570 nm in the yellow spectrum range.

Cyanine3 phosphoramidite 5’ is used in oligonucleotide synthesis for the production of 5’-cyanine3-labeled oligonucleotides. The reagent is compatible with various oligonucleotide synthesizers.

This phosphoramidite can be used for the synthesis of fluorescence-labeled primers and hybridization probes such as TaqMan and Molecular Beacon. Such labeled probes can be detected in multiplex real-time PCR in the TAMRA channel.

Condensation: 3 min. Use 0.02 M iodine solution at the oxidation step to avoid degradation of the cyanine dye.

Deprotection: At room temperature with 30% aqueous ammonium solution. It is recommended to use nucleic bases with labile protective groups for deprotection for not more than 2 h at less than 55°C. AMA (30% aqueous ammonium solution/40% aqueous methylamine 1:1 (v/v)) can be used for 10 min at 65°C in the presence of acetyldeoxycytidine. If deoxyguanidine with a dimethylformamidine protective group is used during synthesis, deprotect with 30% aqueous ammonium solution for 2 h at 65°C. If deoxyguanidine with an isobutyryl protective group is used during synthesis, deprotect for 24-36 h at room temperature.