Journals / Harran Tarım ve Gıda Bilimleri Dergisi / 2010 / Cilt: 14 - Sayı: 2

ANALYSIS OF DNA DAMAGE IN ORGANIZMS VIA “SINGLE CELL GEL ELECTROPHORESIS” (TECHNICAL NOTE): COMET ASSAY METHOD

CANLILARDA “TEK HÜCRE JEL ELEKTROFOREZ” YÖNTEMİ İLE DNA HASAR ANALİZİ (TEKNİK NOT): COMET ANALİZ YÖNTEMİ

Pages
77–89
Publication Date
2014-02-21
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

The method is commonly applied in all living cells.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Single cell gel electrophoresis known also as the comet assay is an important method which enables to measure DNA damage and the evaluation of mechanisms of genotoxic and cytotoxiceffects of physical and chemical agents on organisms.
📊
Key Findings & Data:The method is reliable, easily applicable and has been widely used in recent years.
💡
Conclusion & Impact:The method is commonly applied in all living cells.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Alkali Tek Hücre Jel Elektroforezi · Comet Analizi · SCGE

Abstract

Single cell gel electrophoresis known also as the comet assay is an important method which enables to measure DNA damage and the evaluation of mechanisms of genotoxic and cytotoxiceffects of physical and chemical agents on organisms. The method is reliable, easily applicable and has been widely used in recent years. This method depends on the isolated DNA of single cells running in agarose gel on a microscopy slide in an electrophoretic condition and the migration of damaged DNA fragments with different polarity and molecular weights and the evaluation of DNA fragments following the use of DNA specific fluorescence dyes under a microscopy. The assay method determines the single or double-strand breaks caused by theagents such as oxidative stress, chemical agents, drugs, heavy metal toxicity and ultraviolet lights with fast, sensitive, reliable and cheap way as well as requiring the little amount of cell samples. The method is commonly applied in all living cells.

Özet

Comet analiz yöntemi olarak da bilinen tek hücre jel elektroforez yöntemi son yıllarda genişleyen uygulama alanı, güvenilirliği ve uygulaması kolay olması bakımından kimyasal vefiziksel etmenlerin canlılar üzerinde yol açtığı genotoksik ve sitotoksik etkilerin bir göstergesi olan DNA hasar seviyelerinin ölçülmesini sağlayan önemli bir metotdur. Bu metot, izole edilen DNA içeren tek hücrelerin DNA’larının lam üzerinde hazırlanan agar jel içerisinde elektroforetik ortamda yürütülmesi ve hasar seviyesine göre göç eden farklı yük ve molekül ağırlıklarına sahip DNA parçalarının DNA spesifik floresan boya ile boyandıktan sonra, floresan mikroskop altında değerlendirilmesi esasına dayanmaktadır. Metot, oksidatif stres, toksik ağır metaller, kimyasal ajanlar, ilaçlar ve ultraviole gibi çeşitli genotoksik ajanların DNA sarmalları üzerinde oluşturduğu tek veya çift zincir kırıklarını doğru, hassas, hızlı, ucuz ve az bir örnek hacmi kullanarak ölçen bir yöntem olup, tüm canlı hücreleri üzerinde yapılan çalışmalarda yaygın olarak kullanılmaktadır.

🔖

Cite This Article

DİKİLİTAŞ, M., KOÇYİĞİT, A. (2014). ANALYSIS OF DNA DAMAGE IN ORGANIZMS VIA “SINGLE CELL GEL ELECTROPHORESIS” (TECHNICAL NOTE): COMET ASSAY METHOD. Harran Tarım ve Gıda Bilimleri Dergisi, Cilt: 14 - Sayı: 2, 77–89. https://ajindex.com/articles/423298

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (12)

  1. Abas, Y., Touil, N., Kirsch-Volders, M., Angenon, G., Jacobs, M. ve Famelaer, I.D.H. 2007. Evaluation of UV damage at DNA level in Nicotiana plumbaginifolia protoplasts using single-cell gel electrophoresis. Plant Cell Tissue Organ Culture, 91: 145-154.
  2. Achary, V. M. M., Jena, S., Panda, K. K. ve Panda, B. B. 2008. Aluminium induced oxidative stress and DNA damage in root cells of Allium cepa L. Ecotoxicology and Environmental Safety, 70: 300–310.
  3. Angelis, K.J., Dusinska, M. ve Collins, A.R. 1999. Single cell gel electrophoresis: detection of DNA damage at different levels of sensitivity. Electrophoresis, 20:2133-2138.
  4. Bhanoori, M., Venkateswerlu, G., 1998. The alkaline single cell gel electrophoresis: a new test for assessing DNA single strand breaks in Neurospora crassa. Mutation Research, 405:29-34.
  5. De Pinto, M.C., Francis, D. ve De Gara, L. 1999. The redox state of the ascorbate- dehydroascorbate pair as a specific sensor of cell division in tobacco BY-2 cells. Protoplasma, 209: 90-97.
  6. Dhawan, A., Bajpayee, M. ve Parmar, D. 2009. Comet assay: a reliable tool for the assessment of DNA damage in different models. Cell Biolology Toxicology, 25:5–32.
  7. Dikilitas, M., Kocyigit, A. ve Yigit, F. 2009. A molecular-based fast method to determine the extent of DNA damages in higher plants and fungi. African Journal of Biotechnology, 8 (14): 3118-3127.
  8. Dikilitas, M. 2003. Effect of salinity & its interactions with Verticillium albo-atrum on the disease development in tomato (Lycopersicon esculentum Mill.) and lucerne (Medicago sativa & M. media) plants. Ph.D. Thesis, University of Wales, Swansea Ecobichon, D.J. 2001. Toxic effects of pesticides. In: Klaassen CD(ed.) Casarett and Doull’s toxicology. The basic science of poisons. McGraw-Hill, New York, pp 763-810.
  9. Gichner, T. ve Plewa, M.J. 1998. Induction of somatic DNA damage as measured by single cell gel electrophoresis and point mutation in leaves of tobacco plants. Mutation Research, 401:143-152.
  10. Gichner, T., Patkova, Z., Szakova, J., Demnerova, K. 2004. Cadmium induces DNA damage in tobacco roots, but no DNA damage, somatic mutations or homologous recombinations in tobacco leaves. Mutation Research, 559:49-57.
  11. Gichner, T., Mukherjee, A. ve Veleminsky, J. 2006. DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase- I. Mutation Research, 605:17-21.
  12. Gichner, T., Znidar, I. ve Szakova, J. 2008a. Evaluati