2-Amino-dA-CE Phosphoramidite - (10-1085)
5'-Dimethoxytrityl-N2,N6-bis(diisobutylaminomethylidene)-2,6-diamino-2'-deoxypurine riboside-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramiditeDescription
The simplest approach to the design of high affinity primers and probes is to substitute A sites with 2-amino-A, since the 2-amino-A-T base pair is equivalent in strength to the G-T base pair. 2-Amino-A also destabilizes A-G wobble mismatches, thus increasing specificity. In 1998, we introduced a 2-amino-dA monomer which exhibits fast and effective deprotection in ammonium hydroxide and it is stabilized to depurination during synthesis. We now recommend the use of 0.5 M CSO in anhydrous acetonitrile (40-4632-xx) for best results with multiple additions of 2-amino-dA. This is because the bis formamidine protected 2-amino-dA leads to significant strand scission when standard iodine oxidation is used during synthesis. For this reason, we have also added Pac-2-Amino-dA, a monomer with optimized protection to meet the following criteria: stable during oligonucleotide synthesis, oxidation, and detritylation; labile towards common deprotection conditions (NH3, AMA, MeNH2); and the nucleobase protecting groups are cleaved under fairly mild conditions.
Details
Usage
- Coupling: Standard coupling time. Add a maximum of 3 times when using 0.02 M I2 oxidation or use 0.5 M CSO in anhydrous ACN (40-4632-xx) and a 3 minute oxidation time for best results with multiple additions.
- Deprotection: Ammonium hydroxide 24h/55°C; AMA 1.5h/65°C or 17h/room temperature.
| Specifications | |
|---|---|
| Diluent | Anhydrous Acetonitrile |
| Recommended Storage | Refrigerated storage, maximum of 2-8°C, dry |
| Stability In Solution | 2-3 days |
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 | ||||
| 10-1085-02 | 0.25 g | .25grams | 2.39 | 66.33 | 39.8 | 24.88 | 18.09 | 13.27 | 3.32 |
| 10-1085-90 | 100 µmol | .105grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-1085-95 | 50 µmol | .052grams | 0.5 | 3.33 | 2 | 1.25 | 0.91 | 0.67 | 0.17 |
Expedite
| Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
|---|---|---|---|---|---|---|---|---|
| Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
| 10-1085-02 | 0.25 g | .25grams | 3.56 | 0.07 | 64.8 | 40.5 | 29.45 | 4.05 |
| 10-1085-90 | 100 µmol | .105grams | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
| 10-1085-95 | 50 µmol | .052grams | 0.75 | 0.07 | 8.6 | 5.38 | 3.91 | 0.54 |
Technical documents
Certificate of analysis
