Pac-2-Amino-dA-CE Phosphoramidite - (10-1585)
5'-Dimethoxytrityl-N2-phenoxyacetyl-N6-di-n-butylaminomethylidene-2,6-diamino-2'-deoxypurine riboside-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
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10-1585
5'-Dimethoxytrityl-N2-phenoxyacetyl-N6-di-n-butylaminomethylidene-2,6-diamino-2'-deoxypurine riboside-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Description
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: No changes needed from standard method recommended by synthesizer manufacturer.
- Deprotection: 30% Ammonium Hydroxide for 4 hours at 65 °C or 8-17 hours at 55 °C. Compatible with 30% Ammonium Hydroxide/40% Methylamine 1:1 (AMA) for 20 minutes at 65 °C.
Specifications | |
---|---|
Diluent | Anhydrous Acetonitrile |
Recommended Storage | Refrigerated storage, maximum of 2-8°C, dry |
Stability In Solution | 1-2 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 | ||||
10-1585-02 | 0.25 g | .25grams | 2.4 | 66.67 | 40 | 25 | 18.18 | 13.33 | 3.33 |
10-1585-90 | 100 µmol | .104grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
10-1585-95 | 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 | ||||
10-1585-02 | 0.25 g | .25grams | 3.58 | 0.07 | 65.2 | 40.75 | 29.64 | 4.08 |
10-1585-90 | 100 µmol | .104grams | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
10-1585-95 | 50 µmol | .052grams | 0.75 | 0.07 | 8.6 | 5.38 | 3.91 | 0.54 |
Technical documents
Certificate of analysis
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