DNA loops after cell lysis resemble chromatin loops in an intact nucleus
| Autor: | V. V. Olefirenko, A. V. Sivolob, K. S. Afanasieva |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
animal structures Lysis Chemistry fungi nucleoids Biochemistry Chromatin Cell biology lcsh:Biochemistry 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology medicine.anatomical_structure comet assay DNA loop domains medicine bacteria lcsh:QD415-436 cell lysis Nucleus DNA |
| Zdroj: | Ukrainian Biochemical Journal, Vol 90, Iss 5, Pp 43-49 (2018) |
| ISSN: | 2409-4943 |
| DOI: | 10.15407/ubj90.05.043 |
| Popis: | The comet assay has proved itself to be not only a method of detection of DNA damages at the level of individual cells but also an approach for investigation of spatial organization of DNA loop domains in nucleoids. Usually, those nucleoids are obtained after cell lysis in high-salt buffer (e. g. 2.5 M NaCl) with a detergent: these conditions ensure the removal of cell membranes and most of the chromatin proteins, while supercoiled DNA loop domains remain untouched. In this work, we tested the comet assay applied to nucleoids obtained in low-salt solution (1 M NaCl). These nucleoids keep most of the histones and thus contain the loops resembling the chromatin loops to a greater extent. It was shown that, despite some quantitative differences, the most general features of the kinetics of DNA exit are about the same for nucleoids obtained in high- and low-salt conditions. It can be concluded that the DNA loops in high-salt nucleoids can be efficiently used to investigate the spatial DNA organization in chromatin. |
| Databáze: | OpenAIRE |
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