SIMA phosphoramidite, 6-isomer
| Cat. # | Quantity | Price | Lead time | Buy this product |
|---|---|---|---|---|
| DE260 | 250 mg | $202 | in stock | |
| EE260 | 500 mg | $340 | in stock | |
| FE260 | 1 g |
$575
|
in stock | |
| HE260 | 10 g | please inquire | in stock |
SIMA (dichloro-diphenyl-fluorescein) is a xanthene dye with spectral characteristics similar to those of HEX but with a higher quantum yield. SIMA has higher stability during deprotection under alkaline conditions, so deprotection can be run with aqueous ammonium hydroxide at higher temperatures or with AMA (1:1 mixture, concentrated aqueous ammonium hydroxide/40% aqueous methylamine) at room temperature for 2 h or 65°C for 10 min. When used for deprotection with aqueous ammonium hydroxide at 55°C overnight, oligonucleotide-bound SIMA does not degrade, while with HEX the fluorophore degrades by at least 10%.
SIMA phosphoramidite is used in oligonucleotide synthesis to produce fluorescently labeled primers and hybridization probes for quantitative PCR.
Recommendations for using the reagent:
Coupling: 3 min.
Deprotection: Standard conditions with 25% ammonium hydroxide; deprotection time depends on the composition of nucleic acids and their protective groups. AMA (1:1 mixture of concentrated aqueous ammonium hydroxide / 40% aqueous methylamine) can be used for 2 hours at room temperature or 10 min at 65°C.
Absorption and emission spectra of SIMA
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General properties
| Appearance: | white powder |
| Mass spec M+ increment: | 757.1 |
| Molecular weight: | 1065.02 |
| CAS number: | 1411797-05-1 |
| Molecular formula: | C58H64N3Cl2O10P |
| Solubility: | Good solubility in acetonitrile and DCM |
| Quality control: | NMR 1H and 31P, HPLC-MS (95%) |
| Storage conditions: | 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: | 531 |
| ε, L⋅mol−1⋅cm−1: | 92300 |
| Emission maximum, nm: | 555 |
| Fluorescence quantum yield: | 0.63 |
| CF260: | 0.57 |
| CF280: | 0.18 |
Oligo synthesis details
| Diluent: | anhydrous acetonitrile |
| Coupling conditions: | 3 minute coupling time recommended |
| Deprotection conditions: | identical to protected nucleobases |
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