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Year 2019, Volume: 2 Issue: 1, 73 - 76, 31.07.2019

Abstract

References

  • 1. Adams M.D., Celniker S.E., Holt R.A., Evans C.A. & Gocayne J.D. (2000). The Genome Sequence of Drosophila melanogaster. Science, New Series, Vol. 287, No.5461:2185-2195.
  • 2. Augustyniak M. Gladysz M. Dziewie M. 2015. The Comet assay in insects—Status, prospects and benefits for science. Mutat. Res.: Rev. Mutat. Res. http://dx.doi.org/10.1016/j.mrrev. 2015.09.001.
  • 3. Carmona E.R., Guecheva T.N., Creus A. & Marco R. (2011). Proposal of an In Vivo Comet Assay Using Haemocytes of Drosophila melanogaster. Environmental and Molecular Mutagenesis. 52:165-169.
  • 4. Collins A.R. (2004). The Comet Assay for DNA Damage and Repair; Principles, Applications, and Limitations. Molecular Biotechnology. 26: 249-261.
  • 5. Dhawan A., Bajpayee M., Pandey A.K. & Parmar D. (2009). Protocol for the single cell gel electrophoresis/comet assay for rapid genotoxicity assessment. Developmental Toxicology Division Industrial Toxicology Research Centre Marg, Lucknow 226001U.P. India, Retrieved September 21, 2013 from http// www.cometassayindia.org / Protocol %20for %20 Comet %20Assay.PDF.
  • 6. Fairbairn D.W., Olive P.L. & O'Neill K.L. (1995). The comet assay: a comprehensive review. Mutation Research. 339:37-59.
  • 7. Gaivão I. & Sierra L.M. (2014). Drosophila comet assay: insights, uses, and future perspectives. Frontiers in Genetics. August 2014 | Volume5 | Article 304:1-8.
  • 8. Gupta S.C., Mishra M., Sharma A., Balaji T.G.R.D., Kumar R., Mishra RK. & Chowdhuri D.K. (2010). Chlorpyrifos induces apoptosis and DNA damage in Drosophila through generation of reactive oxygen species. Ecotoxicology and Environmental Safety. 73:1415–1423.
  • 9. Langie S.A.S., Azqueta A. & Collins A.R. (2015). The comet assay: past, present, and future. Frontiers in Genetics|www.frontiersin.org 1 August 2015|Volume6|Article266:1-3.
  • 10. Olive P.L. & Banáth J.P. (2006). The comet assay: a method to measure DNA damage in individual cells. Nature Protocols | VOL.1 NO.1: 23-29.
  • 11. Ong C., Yung L.Y.L., Cai Y., Bay B.H. & Baeg G.H. (2014). Drosophila melanogaster as a model organism to study nanotoxicity. Nanotoxicology. ISSN: 1743-5390 (print), 1743-5404 (electronic) Nanotoxicology, Early Online: 1–8. DOI: 10.3109/17435390.2014.940405.
  • 12. Piper M.D.W. & Partridge L. (2018). Drosophila as a model for ageing. BBA - Molecular Basis of Disease. 1864:2707–2717.
  • 13. Sezginer H. &Dane F. (2016). Genotoxic Analysis Methods of Toxic Substances. Turkish Journal of Scientific Compilations. 9 (1): 50-55.
  • 14. Sayed A.E.D.H., Watanabe-Asaka T., Oda S., Kashiwada S.& Mitani H. (2018). γ-H2AX foci as indication for the DNA damage in erythrocytes of medaka (Oryzias latipes) intoxicated with 4-nonylphenol. Environmental Science and Pollution Research. https://doi.org/10.1007/s11356-018-2985-z.
  • 15. Tice R.R., Agurell E., Anderson D., Burlinson B., Hartmann A., Kobayashi H., Miyamae Y., Rojas E., Ryu J-C. & Sasaki Y.F. (2000). Single Cell Gel/Comet Assay: Guidelines for in vitro and in vivo Genetic Toxicology Testing. Environmental and Molecular Mutagenesis. 35:206-221.
  • 16. Tiwari A.K., Pragya P., Ram K.R. & Chowdhuri D.K. (2011). Environmental chemical mediated male reproductive toxicity: Drosophila melanogaster as an alternate animal model. Theriogenology 76:197–216.

AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER

Year 2019, Volume: 2 Issue: 1, 73 - 76, 31.07.2019

Abstract

Genotoxins can cause various disorders in chromosome and DNA structure
such as gene mutations, chromosome abnormalities and DNA chain breaks.  This situation has been associated with the
concept of genetic toxicity. 
Genotoxicity tests have been developed to predict the possible effects
of genotoxins on all living things in nature. 
The single cell gel electrophoresis assay (SCGE, also known as comet
assay) is used as the fastest, easy and sensitive. The Comet method is based on
distinctive migration characteristics of DNA molecules having different
electrical charges and molecular weights at alkaline pH. According to this
method, nucleolus’ and cells can be visualized by staining with a fluorescent
dye by first placing between the agarose layers and then running at alkaline
electrophoresis buffer and lysis solution and lastly applying neutralization
procedure.
The displayed DNA fractures are measured and evaluated according to
tail length. DNA images are graded according to the degree of damage. DNAs that
are not damaged are evaluated as arbitrary units (AU) by scoring 0 and damage
DNAs are scored from 1 to 4 according to the degree of damage. Comet assay is a
useful method in evaluating the potential toxic effect.  Drosophila melanogaster is the most
important non-mammalian organism used in the fields of medicine and biology. Drosophila melanogaster has been used
for many years as an in vivo model organism for potential toxicity and
genotoxicity studies. It is important to determine the damage of genotoxins on
DNA. It was preferred because it was easy to obtain and grow in the laboratory,
the larvae were cheap and the life cycle was short-lived. Comet analysis using Drosophila
melanogaster
tissues to determine the damage of various genotoxins on DNA
is an effective and easy method. This study aims to explain the alkaline Comet
assay aimed to determine the effect of general various genotoxins on Drosophila melanogaster tissues. 

References

  • 1. Adams M.D., Celniker S.E., Holt R.A., Evans C.A. & Gocayne J.D. (2000). The Genome Sequence of Drosophila melanogaster. Science, New Series, Vol. 287, No.5461:2185-2195.
  • 2. Augustyniak M. Gladysz M. Dziewie M. 2015. The Comet assay in insects—Status, prospects and benefits for science. Mutat. Res.: Rev. Mutat. Res. http://dx.doi.org/10.1016/j.mrrev. 2015.09.001.
  • 3. Carmona E.R., Guecheva T.N., Creus A. & Marco R. (2011). Proposal of an In Vivo Comet Assay Using Haemocytes of Drosophila melanogaster. Environmental and Molecular Mutagenesis. 52:165-169.
  • 4. Collins A.R. (2004). The Comet Assay for DNA Damage and Repair; Principles, Applications, and Limitations. Molecular Biotechnology. 26: 249-261.
  • 5. Dhawan A., Bajpayee M., Pandey A.K. & Parmar D. (2009). Protocol for the single cell gel electrophoresis/comet assay for rapid genotoxicity assessment. Developmental Toxicology Division Industrial Toxicology Research Centre Marg, Lucknow 226001U.P. India, Retrieved September 21, 2013 from http// www.cometassayindia.org / Protocol %20for %20 Comet %20Assay.PDF.
  • 6. Fairbairn D.W., Olive P.L. & O'Neill K.L. (1995). The comet assay: a comprehensive review. Mutation Research. 339:37-59.
  • 7. Gaivão I. & Sierra L.M. (2014). Drosophila comet assay: insights, uses, and future perspectives. Frontiers in Genetics. August 2014 | Volume5 | Article 304:1-8.
  • 8. Gupta S.C., Mishra M., Sharma A., Balaji T.G.R.D., Kumar R., Mishra RK. & Chowdhuri D.K. (2010). Chlorpyrifos induces apoptosis and DNA damage in Drosophila through generation of reactive oxygen species. Ecotoxicology and Environmental Safety. 73:1415–1423.
  • 9. Langie S.A.S., Azqueta A. & Collins A.R. (2015). The comet assay: past, present, and future. Frontiers in Genetics|www.frontiersin.org 1 August 2015|Volume6|Article266:1-3.
  • 10. Olive P.L. & Banáth J.P. (2006). The comet assay: a method to measure DNA damage in individual cells. Nature Protocols | VOL.1 NO.1: 23-29.
  • 11. Ong C., Yung L.Y.L., Cai Y., Bay B.H. & Baeg G.H. (2014). Drosophila melanogaster as a model organism to study nanotoxicity. Nanotoxicology. ISSN: 1743-5390 (print), 1743-5404 (electronic) Nanotoxicology, Early Online: 1–8. DOI: 10.3109/17435390.2014.940405.
  • 12. Piper M.D.W. & Partridge L. (2018). Drosophila as a model for ageing. BBA - Molecular Basis of Disease. 1864:2707–2717.
  • 13. Sezginer H. &Dane F. (2016). Genotoxic Analysis Methods of Toxic Substances. Turkish Journal of Scientific Compilations. 9 (1): 50-55.
  • 14. Sayed A.E.D.H., Watanabe-Asaka T., Oda S., Kashiwada S.& Mitani H. (2018). γ-H2AX foci as indication for the DNA damage in erythrocytes of medaka (Oryzias latipes) intoxicated with 4-nonylphenol. Environmental Science and Pollution Research. https://doi.org/10.1007/s11356-018-2985-z.
  • 15. Tice R.R., Agurell E., Anderson D., Burlinson B., Hartmann A., Kobayashi H., Miyamae Y., Rojas E., Ryu J-C. & Sasaki Y.F. (2000). Single Cell Gel/Comet Assay: Guidelines for in vitro and in vivo Genetic Toxicology Testing. Environmental and Molecular Mutagenesis. 35:206-221.
  • 16. Tiwari A.K., Pragya P., Ram K.R. & Chowdhuri D.K. (2011). Environmental chemical mediated male reproductive toxicity: Drosophila melanogaster as an alternate animal model. Theriogenology 76:197–216.
There are 16 citations in total.

Details

Primary Language English
Subjects Structural Biology
Journal Section Articles
Authors

Fahriye Zemheri Navruz 0000-0003-1744-1091

Publication Date July 31, 2019
Published in Issue Year 2019 Volume: 2 Issue: 1

Cite

APA Zemheri Navruz, F. (2019). AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER. Bartın University International Journal of Natural and Applied Sciences, 2(1), 73-76.
AMA Zemheri Navruz F. AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER. JONAS. July 2019;2(1):73-76.
Chicago Zemheri Navruz, Fahriye. “AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER”. Bartın University International Journal of Natural and Applied Sciences 2, no. 1 (July 2019): 73-76.
EndNote Zemheri Navruz F (July 1, 2019) AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER. Bartın University International Journal of Natural and Applied Sciences 2 1 73–76.
IEEE F. Zemheri Navruz, “AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER”, JONAS, vol. 2, no. 1, pp. 73–76, 2019.
ISNAD Zemheri Navruz, Fahriye. “AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER”. Bartın University International Journal of Natural and Applied Sciences 2/1 (July 2019), 73-76.
JAMA Zemheri Navruz F. AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER. JONAS. 2019;2:73–76.
MLA Zemheri Navruz, Fahriye. “AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER”. Bartın University International Journal of Natural and Applied Sciences, vol. 2, no. 1, 2019, pp. 73-76.
Vancouver Zemheri Navruz F. AN OPTIMIZED COMET ASSAY PROTOCOL FOR DROSOPHILA MELANOGASTER. JONAS. 2019;2(1):73-6.