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A panel of colorimetric assays to measure enzymatic activity in the base excision DNA repair pathway

Healing, Eleanor, Charlier, Clara. F, Meira, Lisie and Elliott, Ruan (2019) A panel of colorimetric assays to measure enzymatic activity in the base excision DNA repair pathway Nucleic Acids Research, 47 (11), e61.

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Abstract

DNA repair is essential for the maintenance of genomic integrity, and evidence suggest that interindividual variation in DNA repair efficiency maycontribute to disease risk. However, robust assays suitable for quantitative determination of DNA repair capacity in large cohort and clinical trials are needed to evaluate these apparent associations fully. We describe here a set of microplate-based oligonucleotide assays for high-throughput, non-radioactiveand quantitative determination of repair enzyme activity at individual steps and over multiple steps of the DNA base excision repair pathway. The assays are highly sensitive: using HepG2 nuclear extract, enzyme activities were quantifiable at concentrationsof 0.0002 to 0.181 g per reaction, depending on the enzyme being measured. Assay coefficients of variation are comparable with other microplate-based assays. The assay format requires no specialist equipment and has the potential to be extended for analysis of a wide range of DNA repair enzyme activities. As such, these assays hold considerable promise for gaining new mechanistic insights into how DNA repair is related to individual genetics, disease status or progression and other environmental factors and investigating whether DNA repair activities can be used a biomarker of disease risk.

Item Type: Article
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :
NameEmailORCID
Healing, Eleanore.healing@surrey.ac.uk
Charlier, Clara. F
Meira, LisieL.Meira@surrey.ac.uk
Elliott, RuanR.M.Elliott@surrey.ac.uk
Date : 14 March 2019
Funders : Biotechnology and Biological Sciences Research Council (BBSRC)
DOI : 10.1093/nar/gkz171
Copyright Disclaimer : The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
Depositing User : Charlene King
Date Deposited : 25 Apr 2019 09:38
Last Modified : 13 Aug 2019 13:27
URI: http://epubs.surrey.ac.uk/id/eprint/851684

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