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Efficient Automated Solid‐Phase Synthesis of DNA and RNA 5′‐Triphosphates
Author(s) -
Sarac Ivo,
Meier Chris
Publication year - 2015
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201502844
Subject(s) - oligonucleotide , rna , oligonucleotide synthesis , pyrophosphate , dna , solid phase synthesis , chemistry , combinatorial chemistry , reagent , dna synthesis , phosphoramidite , biochemistry , organic chemistry , gene , enzyme , peptide
A fast, high‐yielding and reliable method for the synthesis of DNA‐ and RNA 5′‐triphosphates is reported. After synthesizing DNA or RNA oligonucleotides by automated oligonucleotide synthesis, 5‐chloro‐saligenyl‐ N , N ‐diisopropylphosphoramidite was coupled to the 5′‐end. Oxidation of the formed 5′‐phosphite using the same oxidizing reagent used in standard oligonucleotide synthesis led to 5′‐ cyclo Sal‐oligonucleotides. Reaction of the support‐bonded 5′‐ cyclo Sal‐oligonucleotide with pyrophosphate yielded the corresponding 5′‐triphosphates. The 5′‐triphosphorylated DNA and RNA oligonucleotides were obtained after cleavage from the support in high purity and excellent yields. The whole reaction sequence was adapted to be used on a standard oligonucleotide synthesizer.