Technical Documents
Description
An alternative protecting scheme for the normal CE phosphoramidites should allow UltraMILD deprotection and should not react with a wider variety of tags and labels. A set of monomers using phenoxyacetyl (Pac) protected dA and 4-isopropyl-phenoxyacetyl (iPr-Pac) protected dG, along with acetyl protected dC, met the desired criteria for UltraMILD deprotection.
We recommend the use of phenoxyacetic anhydride (Pac2O) in Cap A. This modification removes the possibility of exchange of the iPr-Pac protecting group on the dG with acetate from the acetic anhydride capping mix. Cleavage and deprotection can be carried out in 2 hours at room temperature with ammonium hydroxide or 4 hours with 0.05M potassium carbonate in methanol.
Details
Usage
- Coupling: No changes needed from standard method recommended by synthesizer manufacturer.To avoid any exchange of the iPr-Pac group on the dG with acetyl, use the UltraMild Cap Mix A (40-4210-xx/ 40-4212-xx).
- Deprotection: UltraMILD deprotection: 0.05M Potassium Carbonate in Methanol, 4 hours at Room Temperature OR 2 hours at Room Temperature in 30% Ammonium Hydroxide
| Specifications | |
|---|---|
| Diluent | Anhydrous Acetonitrile |
| Storage | Freezer storage, -10 to -30°C, dry |
| Stability | 24 hours |
Dilution/Coupling Data
The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.
ABI 392/394
| Catalog # | Pack Size | Grams/Pack | 0.1M Dil. (mL) | Approximate Number of Additions | |||||
|---|---|---|---|---|---|---|---|---|---|
| LV40 | LV200 | 40nm | 0.2μm | 1μm | 10μm | ||||
| 10-1601-02 | 0.25 g | .25grams | 2.82 | 80.67 | 48.4 | 30.25 | 22 | 16.13 | 4.03 |
| 10-1601-05 | 0.5 g | .5grams | 5.63 | 174.33 | 104.6 | 65.38 | 47.55 | 34.87 | 8.72 |
| 10-1601-10 | 1.0 g | 1grams | 11.26 | 362 | 217.2 | 135.75 | 98.73 | 72.4 | 18.1 |
Expedite
| Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
|---|---|---|---|---|---|---|---|---|
| Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
| 10-1601-02 | 0.25 g | .25grams | 4.2 | 0.07 | 77.6 | 48.5 | 35.27 | 4.85 |
| 10-1601-05 | 0.5 g | .5grams | 8.4 | 0.07 | 161.6 | 101 | 73.45 | 10.1 |
| 10-1601-10 | 1.0 g | 1grams | 16.81 | 0.07 | 329.8 | 206.13 | 149.91 | 20.61 |
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