Liquid chromatography-mass spectrometry analysis of DNA polymerase reaction products.

Chowdhury G, Guengerich FP
Curr Protoc Nucleic Acid Chem. 2011 Chapter 7: Unit 7.16.1-11

PMID: 22147421 · PMCID: PMC3678528 · DOI:10.1002/0471142700.nc0716s47

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.

© 2011 by John Wiley & Sons, Inc.

MeSH Terms (8)

Base Sequence Chromatography, Liquid DNA-Directed DNA Polymerase DNA Primers Oligonucleotides Sequence Analysis, DNA Tandem Mass Spectrometry Uracil-DNA Glycosidase

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