N6-Methyl-A-CE Phosphoramidite

default
10-3005
5'-Dimethoxytrityl-N-methyl-N-phenoxyacetyl-Adenosine,2'-O-TBDMS-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite

Product Specifications

Formula:
C55H70N7O9PSi
M.W.:
1032.25
F.W.:
343.23
Catalog Number: Select Pack Size and Format to Reveal

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
Storage Freezer storage, -10 to -30°C, dry
Stability 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
10-3005-02 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

References

REFERENCE(S)| 1 Y. Fu, D. Dominissini, G. Rechavi, and C. He, Nat Rev Genet, 2014, 15, 293-306.