FAM-dT phosphoramidite

Cat. # Quantity Price Lead time
13460 100 mg $150 in stock
23460 250 mg $275 in stock
43460 1 g $950 in stock
63460 5 g $3830 in stock
83460 10 g please inquire in stock
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This phosphoramidite allows fluorescein (FAM) to be introduced into any position in the oligonucleotide sequence during synthesis by the phosphoramidite method (in the middle of the chain, at the 5’- and 3’-terminus). The reagent is a conjugate of deoxythymidine phosphoramidite and 6-isomer of FAM. Modification is performed during oligonucleotide synthesis by substituting standard dT phosphoramidite with fluorescein-dT phosphoramidite. This modification does not affect exonuclease or polymerase activity.

For modification with fluorescein at the 5’-terminus use FAM phosphoramidite, 6-isomer.

Usage

Coupling: 10 minutes

Deprotection: standard conditions using ammonium hydroxide; deprotection time depends on oligonucleotide composition and nucleobase protecting groups (deprotection for 17 h at 55 °С 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% of 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 (e.g. 10 min at 65 °C).

Absorption and emission spectra of FAM

Absorption and emission spectra of FAM

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General properties

Appearance: off white powder
Molecular weight: 1423.54
Molecular formula: C79H87N6O17P
Solubility: good in acetonitrile, DCM
Quality control: NMR 1H, NMR 31P, HPLC-MS
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.
MSDS: Download
Product specifications

Spectral properties

Excitation/absorption maximum, nm: 492
ε, L⋅mol−1⋅cm−1: 74000
Emission maximum, nm: 517
Fluorescence quantum yield: 0.93
CF260: 0.22
CF280: 0.17

Oligo synthesis details

Diluent: acetonitrile
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