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Title: | The enzyme-modified comet assay: Enzyme incubation step in 2 vs 12-gels/slide systems | Authors: | Muruzabal, Damian LANGIE, Sabine Pourrut, Bertrand Azqueta, Amaya |
Issue Date: | 2019 | Publisher: | ELSEVIER | Source: | MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS, 845(SI) (Art N° UNSP 402981) | Abstract: | The enzyme-modified comet assay is a commonly used method to detect specific DNA lesions. However, still a lot of errors are made by many users, leading to dubious results and even misinterpretations. This technical note describes some critical points in the use of the enzyme-modified comet assay, such as the enzyme concentration, the time of incubation, the format used and the equipment. To illustrate the importance of these conditions/ parameters, titration experiments of formamidopyrimidine DNA glycosylase (Fpg) were performed using the 2 gels/slide and the 12 minigels/slide formats (plus the 12-Gel Comet Assay Unit (TM)). Incubation times of 15 and 30 min, and 1 h were used. Results showed that the 12 minigels/slide system requires a lower volume and concentration of Fpg. A longer time of incubation has a bigger impact when using such format. Moreover, the paper describes how to perform and interpret a titration experiment when using the enzyme. modified comet assay. | Notes: | [Muruzabal, Damian; Azqueta, Amaya] Univ Navarra, Dept Pharmacol & Toxicol, C Irunlarrea 1,CIFA Bldg, Pamplona 31009, Spain. [Langie, Sabine A. S.] VITO Sustainable Hlth, Boeretang 200, B-2400 Mol, Belgium. [Langie, Sabine A. S.] Hasselt Univ, Ctr Environm Sci, Diepenbeek, Belgium. [Pourrut, Bertrand] Univ Lille Nord France, ISA Lille LGCgE, 48 Blvd Vauban, F-59046 Lille, France. [Azqueta, Amaya] Navarra Inst Hlth Res, IdiSNA, Navarra, Spain. | Keywords: | Comet assay; Formamidopyrimidine DNA glycosylase; Titration; 12 minigels; 2 gels; Enzyme incubation;Comet assay; Formamidopyrimidine DNA glycosylase; Titration; 12 minigels; 2 gels; Enzyme incubation | Document URI: | http://hdl.handle.net/1942/29985 | ISSN: | 1383-5718 | e-ISSN: | 1879-3592 | DOI: | 10.1016/j.mrgentox.2018.11.005 | ISI #: | 000489193400004 | Rights: | 2018 Elsevier B.V. All rights reserved.T | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2020 |
Appears in Collections: | Research publications |
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muruzabal 1.pdf Restricted Access | Published version | 1.15 MB | Adobe PDF | View/Open Request a copy |
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