Unit

UNIT 7.16 Liquid Chromatography-Mass Spectrometry Analysis of DNA Polymerase Reaction Products

  1. Goutam Chowdhury,
  2. F. Peter Guengerich

Published Online: 1 DEC 2011

DOI: 10.1002/0471142700.nc0716s47

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Chowdhury, G. and Guengerich, F. P. 2011. Liquid Chromatography-Mass Spectrometry Analysis of DNA Polymerase Reaction Products. Current Protocols in Nucleic Acid Chemistry. 47:7.16:7.16.1–7.16.11.

Author Information

  1. Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee

Publication History

  1. Published Online: 1 DEC 2011
  2. Published Print: DEC 2011

Abstract

This unit describes experimental and analytical procedures for characterizing the efficiency and fidelity of translesion DNA synthesis across various DNA damages by DNA polymerases in vitro. This procedure utilizes primer extension assays followed by LC-MS and LC-MS/MS analysis of the extension products. Detailed explanations for the analysis of the LC-MS/MS data for deciphering the nucleotide sequences of the DNA fragments are also presented. This approach provides a significant improvement over conventional methods, as it allows detection of misincorporation, as well as frameshift products. Curr. Protoc. Nucleic Acid Chem. 47:7.16.1-7.16.11. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • DNA polymerase;
  • translesion synthesis;
  • sequencing;
  • LC-MS