2'-F-G-CE Phosphoramidite
Product Specifications
2'-Deoxy-2'-fluoro-nucleosides
adopt an RNA-type sugar conformation, presumably due to the high
electronegativity of fluorine. Because of this sugar conformation, RNA
duplexes (A-form) are generally more thermodynamically stable than DNA
duplexes (B-form). As expected, the addition of 2'-F-RNA residues to
oligodeoxynucleotides progressively increases the thermal stability of
their duplexes with RNA. The stabilization is additive at approximately
2° per residue. This compares favorably with 2'-OMe-RNA at around 1.5°
and RNA at 1.1° per residue. In the meantime, base pair specificity
remains intact.
2'-F-RNA phosphodiester linkages are not nuclease resistant, although
the corresponding phosphorothioate linkages are highly resistant.
Researchers usually design antisense oligonucleotides to form duplexes
with RNA, which are then substrates for RNase H. Uniformly modified
2'-F-RNA/RNA duplexes are not substrates for RNase H. However, it is
straightforward to prepare chimeric 2'-F-RNA/DNA phosphorothioate
oligonucleotides which exhibit enhanced binding to the RNA target, are
substrates for RNase H, and are highly nuclease resistant.
REFERENCE(S)
1 E. Viazovkina, M.M. Mangos, M.I. Elzagheid, and M.J. Damha, Curr Protoc Nucleic Acid Chem, 2002, Chapter 4, Unit 4 15.
2 J.K. Watts, and M.J. Damha, Can. J. Chem., 2008, 86, 641-656.
3 J.K. Watts, A. Katolik, J. Viladoms, and M.J. Damha, Org Biomol Chem, 2009, 7, 1904-10.
4 A. Kalota, et al., Nucleic Acids Res., 2006, 34, 451.
5 G.F. Deleavey, et al., Nucleic Acids Res., 2010, 38, 4547-4557,
J.K. Watts, et al., Nucleic Acids Res., 2007, 35, 1441-1451, T. Dowler,
et al., Nucleic Acids Res., 2006, 34, 1669-1675.
Details
Usage
- Coupling: 3 minute coupling time recommended.
- Deprotection:
Ammonium Hydroxide for 17 hours at 55°C or 30% Ammonia Hydroxide/40%
Methylamine 1:1 (AMA) for 2 hours at Room Temperature. (Note - heating
in AMA will lead to some degradation of the 2'-Fluoro nucleotides.)
| Specifications |
|---|
| Diluent | Anhydrous Acetonitrile |
| Recommended Storage | Freezer storage, -10 to -30°C, dry |
| Stability In Solution | 2-3 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 |
|---|
| 10-3420-02 | 0.25 g | .25grams | 2.91 | 83.67 | 50.2 | 31.38 | 22.82 | 16.73 | 4.18 |
| 10-3420-05 | 0.5 g | .5grams | 5.83 | 181 | 108.6 | 67.88 | 49.36 | 36.2 | 9.05 |
Expedite
| Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions |
|---|
| Molarity | 50nm | 0.2μm | 1μm | 15μm |
|---|
| 10-3420-02 | 0.25 g | .25grams | 4.35 | 0.07 | 80.6 | 50.38 | 36.64 | 5.04 |
| 10-3420-05 | 0.5 g | .5grams | 8.7 | 0.07 | 167.6 | 104.75 | 76.18 | 10.48 |
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