Enzymatic Synthesis of Backbone‐Modified Oligonucleotides Using T4 DNA Ligase. Issue 2 (28th January 2019)

• Record Type: Journal Article
• Authors: Kestemont, Donaat; Herdewijn, Piet; Renders, Marleen
• Editors: Arkin, Adam P.; Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 11:Issue 2(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-01-28
• Subjects: backbone‐modified oligonucleotides -- crowding agents -- enzymatic synthesis -- mixed‐chemistry -- T4 DNA ligase; Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/cpch.62
• Languages: English
• ISSNs: 2160-4762

Abstract

Abstract: T4 DNA ligase in high concentrations of certain crowding agents and cosolutes catalyzes the synthesis of a series of backbone‐modified oligonucleotides that are difficult to obtain chemically. Backbone‐modified nucleic acids are often enzymatically and chemically more stable, making them interesting as potential diagnostic or therapeutic agents, as a biosafety tool, or in nanotechnology. In this article, we describe a small‐scale experiment to probe the efficiency of the ligation reaction of modified oligonucleotides in the presence of 3 M betaine and 10% PEG 8000, followed by large‐scale ligation with subsequent isolation of the ligated oligonucleotide. The correct product formation can be verified using denaturing polyacrylamide gel electrophoresis and mass spectrometry. © 2019 by John Wiley & Sons, Inc.