HEX phosphoramidite, 6-isomer
| Cat. # | Quantity | Price | Lead time | Buy this product |
|---|---|---|---|---|
|
CB260
/ Easily soluble lyophilised form in 6 mL septum
|
100 mg | – | in stock | |
|
DB260
/ Easily soluble lyophilised form in 6 mL septum
|
250 mg | $272 | in stock | |
|
FB260
/ Easily soluble lyophilised form in 20 mL septum
|
1 g |
$575
|
in stock | |
| GB260 | 5 g |
$2640
|
in stock | |
| HB260 | 10 g | please inquire | in stock |
HEX phosphoramidite for oligonucleotide synthesis, pure 6-isomer.
HEX (hexachlorofluorescein) is a fluorescein derivative with emission in the yellow spectrum range (absorption maximum at 533 nm, emission maximum at 549 nm).
HEX phosphoramidite is used for synthesis of fluorescent-labeled primers and hybridization probes such as TaqMan, Molecular Beacon, and Scorpion for qPCR. HEX is most effectively quenched by non-fluorescent DusQ 1 dark quencher because of significant overlapping of their spectra (convenient for use with DusQ 1 CPG 500 solid support with a pore size of 500 Å). Many automated sequencers based on capillary gel electrophoresis have a detection channel for HEX. Therefore, this phosphoramidite is commonly used for synthesis of 5’-labeled oligonucleotides for fragment analysis, particularly for microsatellite analysis, when microsatellite loci are amplified using a fluorescent-labeled forward primer and a non-labeled reverse primer.
Usage
Coupling: 3 min.
Deprotection: standard conditions using 25% ammonium; 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 concentrated aqueous ammonium/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 HEX
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| Appearance: | off white solid |
| Molecular weight: | 1050.61 |
| CAS number: | 1360547-55-2 |
| Molecular formula: | C46H52N3Cl6O10P |
| Solubility: | Good solubility in acetonitrile and DCM |
| Quality control: | NMR 1H and HPLC-MS (95+%) |
| 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: | 533 |
| ε, L⋅mol−1⋅cm−1: | 87770 |
| Emission maximum, nm: | 549 |
| Fluorescence quantum yield: | 0.57 |
| CF260: | 0.30 |
| CF280: | 0.13 |
Oligo synthesis details
| Diluent: | anhydrous acetonitrile (prepare a 0.1 М solution, storage 1 week). |
| Coupling conditions: | standard coupling, identical to normal nucleobases |
| Deprotection conditions: | identical to protected nucleobases |
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