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Int. J. Mol. Sci. 2014, 15(10), 18023-18039; doi:10.3390/ijms151018023

Optimization of a DNA Nicking Assay to Evaluate Oenocarpus bataua and Camellia sinensis Antioxidant Capacity

Université des Antilles et de la Guyane, UMR QUALITROP, campus universitaire de Troubiran, P.O. Box 792, 97337 Cayenne Cedex, French Guiana, France
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Received: 31 July 2014 / Revised: 18 September 2014 / Accepted: 25 September 2014 / Published: 9 October 2014
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

This study was aimed at assessing the DNA damage protective activity of different types of extracts (aqueous, methanolic and acetonic) using an in vitro DNA nicking assay. Several parameters were optimized using the pUC18 plasmid, especially FeSO4, EDTA, solvent concentrations and incubation time. Special attention has been paid to removing the protective and damaging effect of the solvent and FeSO4 respectively, as well as to identifying the relevant positive and negative controls. For each solvent, the optimal conditions were determined: (i) for aqueous extracts, 0.33 mM of FeSO4 and 0.62 mM of EDTA were incubated for 20 min at 37 °C; (ii) for acetone extracts, 1.16% solvent were incubated for 15 min at 37 °C with 1.3 mM of FeSO4 and 2.5 mM of EDTA and (iii) for methanol extracts, 0.16% solvent, were incubated for 1.5 h at 37 °C with 0.33 mM of FeSO4 and 0.62 mM of EDTA. Using the optimized conditions, the DNA damage protective activity of aqueous, methanolic and acetonic extracts of an Amazonian palm berry (Oenocarpus bataua) and green tea (Camellia sinensis) was assessed. Aqueous and acetonic Oenocarpus bataua extracts were protective against DNA damage, whereas aqueous, methanolic and acetonic extracts of Camellia sinensis extracts induced DNA damage. View Full-Text
Keywords: pUC18; antioxidant; prooxidant; DNA nicking assay; Fenton; Amazonian palm; Oenocarpus bataua; Camellia sinensis pUC18; antioxidant; prooxidant; DNA nicking assay; Fenton; Amazonian palm; Oenocarpus bataua; Camellia sinensis
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Leba, L.-J.; Brunschwig, C.; Saout, M.; Martial, K.; Vulcain, E.; Bereau, D.; Robinson, J.-C. Optimization of a DNA Nicking Assay to Evaluate Oenocarpus bataua and Camellia sinensis Antioxidant Capacity. Int. J. Mol. Sci. 2014, 15, 18023-18039.

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