Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation
Reference materials are needed to quantify the level of DNA damage in cells, to assess sources of measurement variability and to compare results from different laboratories. The comet assay (single cell gel electrophoresis) is a widely used method to determine DNA damage in the form of strand breaks...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2020-01-01
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| Series: | Journal of Nucleic Acids |
| Online Access: | http://dx.doi.org/10.1155/2020/2928104 |
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| author | Donald H. Atha Omobola Cole Breece Clancy Alessandro Tona Vytas Reipa |
| author_facet | Donald H. Atha Omobola Cole Breece Clancy Alessandro Tona Vytas Reipa |
| author_sort | Donald H. Atha |
| collection | DOAJ |
| description | Reference materials are needed to quantify the level of DNA damage in cells, to assess sources of measurement variability and to compare results from different laboratories. The comet assay (single cell gel electrophoresis) is a widely used method to determine DNA damage in the form of strand breaks. Here we examine the use of electrochemical oxidation to produce DNA damage in cultured mammalian cells and quantify its percentage using the comet assay. Chinese hamster ovary (CHO) cells were grown on an indium tin oxide electrode surface and exposed 12 h to electrochemical potentials ranging from 0.5 V to 1.5 V (vs Ag/AgCl). The resulting cells were harvested and analyzed by comet and a cell viability assay. We observed a linear increase in the percentage (DNA in tail) of strand breaks along with a loss of cell viability with increasing oxidation potential value. The results indicate that electrochemically induced DNA damage can be produced in mammalian cells under well-controlled conditions and could be considered in making a cellular reference material for the comet assay. |
| format | Article |
| id | doaj-art-8a21cc771699430d80e03f75f836bb87 |
| institution | Kabale University |
| issn | 2090-0201 2090-021X |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nucleic Acids |
| spelling | doaj-art-8a21cc771699430d80e03f75f836bb872025-02-03T06:46:37ZengWileyJournal of Nucleic Acids2090-02012090-021X2020-01-01202010.1155/2020/29281042928104Cellular Reference Materials for DNA Damage Using Electrochemical OxidationDonald H. Atha0Omobola Cole1Breece Clancy2Alessandro Tona3Vytas Reipa4Materials Measurement Laboratory, Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USAMaterials Measurement Laboratory, Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USAMaterials Measurement Laboratory, Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USAMaterials Measurement Laboratory, Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USAMaterials Measurement Laboratory, Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USAReference materials are needed to quantify the level of DNA damage in cells, to assess sources of measurement variability and to compare results from different laboratories. The comet assay (single cell gel electrophoresis) is a widely used method to determine DNA damage in the form of strand breaks. Here we examine the use of electrochemical oxidation to produce DNA damage in cultured mammalian cells and quantify its percentage using the comet assay. Chinese hamster ovary (CHO) cells were grown on an indium tin oxide electrode surface and exposed 12 h to electrochemical potentials ranging from 0.5 V to 1.5 V (vs Ag/AgCl). The resulting cells were harvested and analyzed by comet and a cell viability assay. We observed a linear increase in the percentage (DNA in tail) of strand breaks along with a loss of cell viability with increasing oxidation potential value. The results indicate that electrochemically induced DNA damage can be produced in mammalian cells under well-controlled conditions and could be considered in making a cellular reference material for the comet assay.http://dx.doi.org/10.1155/2020/2928104 |
| spellingShingle | Donald H. Atha Omobola Cole Breece Clancy Alessandro Tona Vytas Reipa Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation Journal of Nucleic Acids |
| title | Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation |
| title_full | Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation |
| title_fullStr | Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation |
| title_full_unstemmed | Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation |
| title_short | Cellular Reference Materials for DNA Damage Using Electrochemical Oxidation |
| title_sort | cellular reference materials for dna damage using electrochemical oxidation |
| url | http://dx.doi.org/10.1155/2020/2928104 |
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