FAM-dT phosphoramidite
| Cat. # | Quantity | Price | Lead time | Buy this product |
|---|---|---|---|---|
| 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 |
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
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R6G NHS ester, 5-isomer
R6G (rhodamine 6G) NHS ester, pure 5-isomer. This dye is often used for the labeling of oligonucleotides for qPCR.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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