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Updated: Jan 16, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Use of a Dihydroxyacetone Derivative as Protecting Reagent to Phosphorylate Oligonucleotides
1Département de Chimie Moléculaire, CNRS UMR 5250, Université Grenoble Alpes, Grenoble, France.
Abstract:
We present here a new reagent enabling the supported synthesis of oligodeoxynucleotides (ODNs) and oligoribonucleotides (ORNs) containing a phosphate group at the 5'-terminal position after complete deprotection. This reagent, derived from a dihydroxyacetone core, contains a dimethoxytrityl (DMTr) group. The procedure for final deprotection is very similar to that routinely used in the synthesis of unmodified ODNs or ORNs. In particular, it preserves the advantages of the "trityl-on" method, namely facile purification by reverse-phase high-performance liquid chromatography (RP-HPLC), short treatment with an acetic acid solution in water, the possibility of on-resin monitoring by trityl cation assay, and the use of equipment commonly founded in chemical laboratories. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Phosphorylation of oligodeoxyribonucleotides (ODNs) Alternate Protocol: Phosphorylation of oligoribonucleotides (ORNs) Support Protocol: Preparation of phosphorylation reagent 1.
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