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TAMRA phosphoramidite, 5-isomer

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 我们建议使用TAMRA 亚磷酰胺,6-异构体 (TAMRA phosphoramidite, 6-isomer)作为替代品
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This phosphoramidite is used for synthesis of oligonucleotides 5’-labeled with TAMRA.

TAMRA (carboxytetramethylrhodamine) is a xanthene dye from the rhodamine family with emission in the orange spectrum range (maximum at 563 nm). This fluorophore is traditionally used as a FRET-acceptor (and a quencher) in a pair with fluorescein (FAM) due to significant overlapping of their spectra. Thus, this phosphoramidite is convenient for the synthesis of dual-labeled probes TaqMan, which contain 5’-terminal TAMRA and FAM in the middle of the sequence or at the 3’-end (using Fluorescein dT Phosphoramidite and FAM CPG, respectively).

TAMRA 5’-labeled oligonucleotides are commonly used for quantitative PCR and fragment analysis (for example, for microsatellite marker analysis) because the equipment available has a detection channel for TAMRA frequently.

The TAMRA dye is not stable in the presence of ammonium and sterically non-hindered primary amines, so it is strongly recommended to follow specified conditions for labeled oligonucleotide deprotection.

Usage

Coupling: 7.5 min.

Deprotection: tret-buthylamine : methanol : water 1 : 1 : 3 (v/v/v) («TAMRA cocktail») for 6 hours at 60 °C, then cool down to room temperature.

Due to complete and irreversible degradation of the TAMRA dye, do NOT use aqueous ammonium and AMA for deprotecting a modified oligonucleotide from the solid-phase support.

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General properties

Appearance: purple solid
Mass spec M+ increment: 589.60
Molecular weight: 727.83
Molecular formula: C40H50N5O6P
Solubility: good in acetonitrile, dichloromethane
Quality control: NMR 1H, HPLC-MS (95%), coupling test
Storage conditions: Storage: 12 months after receival at -20°C in the dark. Transportation: at room temperature for up to 3 weeks. Avoid prolonged exposure to light. Desiccate.
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产品规格

Spectral properties

Excitation/absorption maximum, nm: 541
ε, L⋅mol−1⋅cm−1: 84000
Emission maximum, nm: 567
CF260: 0.32
CF280: 0.19

Oligo synthesis details

Diluent: acetonitrile
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