UNIT 4.67 Solid‐Phase Synthesis of DNA and RNA 5′‐O‐Triphosphates Using cycloSal Chemistry. Issue 1 (1st March 2016)

• Record Type: Journal Article
• Authors: Sarac, Ivo; Meier, Chris
• Is Part Of: Current protocols in nucleic acid chemistry. Volume 64:Issue 1(2016)
• Page Start: 4.67.1
• Page End: 4.67.13
• Publication Date: 2016-03-01
• Subjects: automated solid‐phase synthesis -- 5′‐O‐triphosphate -- cycloSal -- modified DNA and RNA synthesis -- modified oligonucleotides; Nucleic acids -- Laboratory manuals; Nucleic Acids -- chemical synthesis; Nucleic acids; Laboratory Manuals; Electronic books; Laboratory manuals; 572.8028
• DOI: 10.1002/0471142700.nc0467s64
• Languages: English
• ISSNs: 1934-9270

Abstract

Abstract : A chemical method for the synthesis of short strands of DNA and RNA 5′‐ O ‐triphosphates is described that makes use of the conventional coupling and oxidizing reagents that are readily available on standard DNA/RNA synthesizers. After automated solid‐phase synthesis of oligonucleotides and 5′‐ O ‐detritylation, a novel cyclo Sal‐phosphoramidite, 5‐chloro‐saligenyl‐ N, N ‐diisopropylphosphoramidite, was coupled to the 5′‐hydroxyl group and the product oxidized. The resulting support‐bonded 5′‐ cyclo Sal‐oligonucleotide was reacted with pyrophosphate to yield 5′‐ O ‐triphosphorylated DNA/RNA oligonucleotides after cleavage from the support in high purity and excellent yields. The reaction sequence was adapted to be used completely on a standard automated oligonucleotide synthesizer. © 2016 by John Wiley & Sons, Inc.