Fluorescein-dT phosphoramidite

Cat. # Quantity Price Lead time
13460 100 mg –   5 days
23460 250 mg $410.00 in stock
43460 1 g $1590.00 in stock
63460 5 g $6900.00 in stock
83460 10 g please inquire in stock

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
MSDS: Download
Product specifications

Spectral properties

Excitation/absorption maximum, nm: 490
ε, L⋅mol−1⋅cm−1: 80000
Emission maximum, nm: 513
Fluorescence quantum yield: 0.93
CF260: 0.20
CF280: 0.17

Oligo synthesis details

Diluent: acetonitrile
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