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Therefore, a method reliably assessing DNA ligation capacity in protein extracts from murine tissues was aimed to establish. To optimize applicability, the use of radioactively labeled substrates was avoided and replaced by fluorescently labeled oligonucleotides. Briefly, tissue extracts were incubated with those complementary oligonucleotides so that in an ensuing gel electrophoresis ligated strands could be separated from unconnected molecules. Originally, the method was intended for use in cerebellum tissue to further elucidate possible mechanisms of neurodegenerative diseases. However, due to its inhomogeneous anatomy, DNA ligation efficiency varied strongly between different cerebellar areas, illuminating the established assay to be suitable only for homogenous organs. Thus, for murine liver tissue sufficient intra\u2010 and interday repeatability was shown during validation. In further experiments, the established assay was applied to an animal study comprising young and old (24 and 110\u2009weeks) mice which showed that DNA ligation efficiency was affected by neither sex nor age. Finally, the impact of in vitro addition of the trace elements copper, iron, and zinc on DNA ligation in tissue extracts was investigated. While all three metals inhibited DNA ligation, variations in their potency became evident. In conclusion, the established method can be reliably used for investigation of DNA ligation efficiency in homogenous murine tissues.<\/jats:p>","DOI":"10.1002\/em.22602","type":"journal-article","created":{"date-parts":[[2024,5,20]],"date-time":"2024-05-20T10:09:55Z","timestamp":1716199795000},"page":"106-115","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of a nonradioactive <scp>DNA<\/scp> ligation assay and its applications in murine tissues"],"prefix":"10.1002","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2916-1740","authenticated-orcid":false,"given":"Sharleen","family":"Friese","sequence":"first","affiliation":[{"name":"Department of Food Chemistry, Institute of Nutritional Science University of Potsdam  Nuthetal Germany"},{"name":"TraceAge \u2013 DFG Research Unit on Interactions of Essential Trace Elements in Healthy and Diseased Elderly (FOR 2558)  Berlin\u2010Potsdam\u2010Jena\u2010Wuppertal Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tom","family":"Heinze","sequence":"additional","affiliation":[{"name":"Department of Food Chemistry, Institute of Nutritional Science University of Potsdam  Nuthetal Germany"},{"name":"TraceAge \u2013 DFG Research Unit on Interactions of Essential Trace Elements in Healthy and Diseased Elderly (FOR 2558)  Berlin\u2010Potsdam\u2010Jena\u2010Wuppertal Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Franziska","family":"Ebert","sequence":"additional","affiliation":[{"name":"Department of Food Chemistry, Institute of Nutritional Science University of Potsdam  Nuthetal Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tanja","family":"Schwerdtle","sequence":"additional","affiliation":[{"name":"Department of Food Chemistry, Institute of Nutritional Science University of Potsdam  Nuthetal Germany"},{"name":"TraceAge \u2013 DFG Research Unit on Interactions of Essential Trace Elements in Healthy and Diseased Elderly (FOR 2558)  Berlin\u2010Potsdam\u2010Jena\u2010Wuppertal Germany"},{"name":"German Federal Institute for Risk Assessment (BfR)  Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,5,20]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0891-5849(99)00091-x"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0378-4274(99)00273-8"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M404524200"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41380-019-0375-7"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0027-5107(02)00003-9"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells9071665"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky1202"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/1878-0261.13285"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-0270(97)00079-4"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0891-5849(01)00483-x"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00775-013-1012-6"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms231911010"},{"key":"e_1_2_10_14_1","unstructured":"European Medicines Agency. 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