TET phosphoramidite for synthesis of fluorescently labeled oligonucleotides, pure 6-isomer.
TET (tetrachlorofluorescein) is a green-fluorescent fluorescein derivate (absorption maximum at 519 nm, emission maximum at 535 nm).
TET phosphoramidite is used for synthesis of fluorescently-labeled primers and hybridization probes for qPCR. TET can be used with DusQ1 fluorescence quencher (can be used with 500 Å DusQ1 CPG 500).
5’-labeled primers are used with non-labeled reverse primers for microsatellite amplification via PCR followed by fragment analysis. TET-labeled amplification products can be analyzed using various sequencers for capillary electrophoresis, including ABI PRISM® 310 Genetic Analyzer.
Condensation: 3 min.
Deprotection: standard conditions with 25% ammonium hydroxide; deprotection time depends on oligonucleotide composition and nucleobase protecting groups (deprotection for 17 hours at 55°C removes all protecting groups from standard nucleobases). AMA (solution of 30% ammonium hydroxide/40% aqueous methylamine 1:1 v/v) can be used with ~5% non-fluorescent side product forming. To avoid formation of the side product, start deprotection with ammonium hydroxide (30 min at room temperature), then add an equal volume of 40% aqueous methylamine and continue deprotection as required with AMA (10 min at 65°C).
