Phosphorylating Reagent for 5'-Triphosphorylation of Oligonucleotides
5-Chlorosaligenyl-N,N-diisopropylphosphoramidite (C3326) is a phosphorylating reagent used to introduce a triphosphate group onto the 5' end of oligonucleotides. It is effective in the triphosphorylation of 5'-hydroxy groups and applicable to both RNA and DNA, and it can be used in an oligonucleotide synthesizer. In the reaction sequence, the 5'-hydroxy group is phosphitylated with C3326 in the presence of BTT in CH3CN, then oxidized with iodine in pyridine/H2O/THF to give a 5'-cyclophosphate ester intermediate. This intermediate then reacts with Bis(tetrabutylammonium) Dihydrogen Pyrophosphate (B5440) in DMF to attach the triphosphate group. The sequence is applicable to both solid-phase synthesis and standard oligonucleotide synthesizers, and final deprotection uses aqueous NH3/aqueous CH3NH2 (AMA), with TBAF (T1338) for RNA.¹)
Related products
- TCI C3326 1620086-77-2 5-Chlorosaligenyl-N,N-diisopropylphosphoramidite
- TCI B5440 857447-79-1 Bis(tetrabutylammonium) Dihydrogen Pyrophosphate
- TCI B3020 21871-47-6 5-(Benzylthio)-1H-tetrazole
- TCI I0604 7553-56-2 Iodine
- TCI T1338 429-41-4 Tetrabutylammonium Fluoride (ca 1mol/L in Tetrahydrofuran)
References
- Solid‐Phase Synthesis of DNA and RNA 5′‐O‐Triphosphates Using cycloSal Chemistry. I. Sarac, C. Meier, Curr. Protoc. Nucleic Acid Chem. 2016, 64:4.67.1-4.67.14. doi:10.1002/0471142700.nc0467s64
Source
- TCI Product Highlights: www.tcichemicals.com/ID/en/product/tci-topics/ProductHighlights_20191125
- TCI product page C3326: tcichemicals.com/ID/en/p/C3326
- TCI product page B5440: tcichemicals.com/ID/en/p/B5440
- TCI brochure PP009: Phosphorylating Reagents for the Synthesis of Nucleotide 5'-O-Triphosphates
For research and laboratory use only, not for human consumption.
