Unit

UNIT 10.6 Base Composition Analysis of Nucleosides Using HPLC

  1. Alex Andrus1,
  2. Robert G. Kuimelis2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142700.nc1006s01

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Andrus, A. and Kuimelis, R. G. 2001. Base Composition Analysis of Nucleosides Using HPLC. Current Protocols in Nucleic Acid Chemistry. 1:10.6:10.6.1–10.6.6.

Author Information

  1. 1

    PE Applied Biosystems, Foster City, California

  2. 2

    Phylos, Inc., Lexington, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 2000

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.