N6-Methyl-A-CE Phosphoramidite
5'-Dimethoxytrityl-N-methyl-N-phenoxyacetyl-Adenosine,2'-O-TBDMS-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Product Specifications
Description
Methylation
of adenosine at position 1 produces a drastic functional change in the
nucleobase. 1-Methyladenosine (pKa 8.25) is a much stronger base than
adenosine (pKa 3.5). N-1 methylation excludes participation of the
adenine base in canonical Watson–Crick base pairing and provides a
positive charge to the nucleobase. This modification also alters the
hydrophobicity of the base, the stacking properties, the ordering of
water molecules and the chelation properties. The base may become
involved in non-canonical hydrogen bonding, in electrostatic
interactions and, in general, it may contribute to the conformational
dynamics of the tRNA.
In the central dogma of molecular biology, genetic information flows
from DNA to RNA and then to protein. Reversible epigenetic modifications
on genomic DNA and histone have been known to substantially regulate
gene expression. On the other hand, there exists more than 100 naturally
occurring chemical modifications in RNA; however, the functions of
these RNA modifications are largely unknown. Whether some of these
modifications in RNA can be reversed and could impact gene expression in
the central dogma was unknown until the recent discovery of
N6-methyladenosine (N6-Me-A) as the first example of reversible RNA
methylation.1 We offer the N6-Me-A RNA monomer with a phenoxyacetyl
protecting group to minimize potential branching. We have shown
N6-Me-A-CE Phosphoramidite to be completely compatible with all popular
RNA synthesis and deprotection methods, from UltraMild to the most
popular procedure using AMA for deprotection.
Details
Usage
- Coupling:
12 minute coupling time is recommended. To avoid any exchange of the
Pac group with acetyl, use the UltraMild Cap Mix A (40-4210-xx/
40-4212-xx).
- Deprotection: RNA Deprotection-see bulletin
| Specifications |
|---|
| Diluent | Anhydrous Acetonitrile |
| Recommended Storage | Freezer storage, -10 to -30°C, dry |
| Stability In Solution | 1-2 days |
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 |
|---|
| 10-3005-02 | 0.25 g | .25grams | 2.42 | 67.33 | 40.4 | 25.25 | 18.36 | 13.47 | 3.37 |
| 10-3005-90 | 100 µmol | .103grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-3005-95 | 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 |
|---|
| 10-3005-02 | 0.25 g | .25grams | 3.61 | 0.07 | 65 | 41 | 13.71 | 4 |
| 10-3005-90 | 100 µmol | .103grams | 1.5 | 0.07 | 23 | 14 | 4 | 1 |
| 10-3005-95 | 50 µmol | .052grams | 0.75 | 0.07 | 8 | 5 | 1 | 0 |
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