Base composition analysis of nucleosides using HPLC

Current Protocols in Nucleic Acid Chemistry
A Andrus, R G Kuimelis

Abstract

In this protocol, nuclease digestion of an oligonucleotide is followed by dephosphorylation and HPLC analysis of the monomers on a reversed-phase C18 column. This method can be used to detect and quantitate a wide variety of nucleobase modifications in oligonucleotides. Integrated areas of the nucleoside chromatogram give precise quantitation of nucleoside composition when the relative extinction coefficient cofactors are applied to the sum of the areas of the four bases. The protocol is also useful for analysis of oligonucleotides containing conjugated moieties and carbohydrate modifications.

Citations

Jul 21, 2010·Journal of Visualized Experiments : JoVE·Markus HafnerThomas Tuschl
Jul 7, 2020·Current Protocols in Nucleic Acid Chemistry·Shiyue FangBhaskar Halami
Dec 16, 2020·Analytical Methods : Advancing Methods and Applications·Steven R StrezsakNicholas J Skizim

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