Chemical Phosphorylation Reagent II

default
10-1901
[3-(4,4'-Dimethoxytrityloxy)-2,2-dicarboxyethyl]propyl-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite
Formula:
C39H51N2O9P
M.W.:
722.82
F.W.:
79.98
Catalog: Make Selections to Reveal

Description

Chemical Phosphorylation Reagent is most commonly used to phosphorylate the 5'-terminus of an oligonucleotide. Although this product is also successful in 3'-phosphorylation, 3'-Phosphate CPG allows direct preparation of oligonucleotides with a 3'-phosphate group. Chemical Phosphorylation Reagent II contains a DMT group on a side chain which is stable to base cleavage and can be left on the oligonucleotide for use in RP purification. The DMT group is later removed with aqueous acid and the side chain is eliminated after brief treatment with aqueous ammonium hydroxide to yield the 5'-phosphate.1 Solid CPR II is similar in performance to CPR II but it is easier to prepare aliquots since it is a powder. Many researchers treat synthesis supports with a hindered base (e.g., diethylamine, diisopropylethylamine, or DBU) post-synthesis to eliminate and remove the cyanoethyl phosphate groups. In this way, the acrylonitrile formed in situ is removed from the support and is not available to alkylate dT residues at the N3 position in the oligos. Since the sulfonylethyl group in 3'-Phosphate CPG is also susceptible to ß-elimination leading to oligo cleavage, this technique is not compatible with 3'-phosphate CPG. Using CPR II CPG, which is base labile but does not support ß-elimination, the cyanoethyl groups can be removed from the oligo prior to cleavage and base deprotection. ABI-style vials and columns are supplied unless otherwise requested.

Details

Usage

  • Coupling: 6 minute coupling time recommended.
  • Deprotection: If the final DMT is removed during synthesis, deprotection under standard conditions will eliminate the CPR II to yield a 5' Phosphate. If the DMT is retained, see Technical Bulletin for removal procedure (http://www.glenresearch.com/Technical/TB_CPR_II.pdf).
Specifications
Diluent Anhydrous Acetonitrile
Storage Refrigerated storage, maximum of 2-8°C, dry
Stability 24 hours


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
10-1901-02 0.25 g .25grams 3.46 102 61.2 38.25 27.82 20.4 5.1
10-1901-90 100 µmol .072grams 1 20 12 7.5 5.45 4 1

Expedite

Catalog # Pack Size Grams/Pack Dilution (mL) Approximate Number of Additions
Molarity 50nm 0.2μm 1μm 15μm
10-1901-02 0.25 g .25grams 5.16 0.07 96.8 60.5 44 6.05
10-1901-90 100 µmol .072grams 1.5 0.07 23.6 14.75 10.73 1.48

References

1 A. Guzaev, , A. Azhayev, and H. Lonnberg, Tetrahedron, 1995, 51, 9375-9384.