Microbial Diversity Assessment in Soil from Dumping Sites: Isolation, DNA Extraction and Electrophoretic Analysis


Authors : Nishant Thalwal

Volume/Issue : Volume 9 - 2024, Issue 7 - July


Google Scholar : https://tinyurl.com/4utwaxh9

Scribd : https://tinyurl.com/2y8jkfjp

DOI : https://doi.org/10.38124/ijisrt/IJISRT24JUL1686

Note : A published paper may take 4-5 working days from the publication date to appear in PlumX Metrics, Semantic Scholar, and ResearchGate.


Abstract : This study aims to investigate the microbial diversity present in soil samples collected from various dumping sites, which are known for their complex and heterogeneous waste environments. The primary objective is to isolate and characterize the microbial communities inhabiting these soils, providing insights into their composition and potential ecological roles. Soil samples were collected by digging 5 cm deep at multiple dumping locations to ensure a representative sampling of the microbial populations. The collected soil samples underwent a serial dilution process, extending up to 10^-9 and 10^-10 dilutions, to facilitate the isolation of individual microbial colonies. These diluted samples were then inoculated onto Standard Potato Dextrose Agar (PDA) media, a nutrient-rich medium conducive to microbial growth. Following an incubation period, distinct bacterial colonies were observed and isolated for further analysis. DNA was extracted from the isolated bacterial colonies using a standard DNA extraction protocol. The quality and purity of the extracted DNA were assessed through gel electrophoresis, a technique that separates DNA fragments based on their size. Remarkably, the electrophoretic analysis revealed clear and distinct DNA bands along with RNA bands, indicating successful extraction of high-quality microbial DNA. Notably, this was achieved without the use of proteinase treatment, which is typically employed to remove protein contaminants from DNA samples. The results of this study underscore the robustness of the methodologies employed in isolating and characterizing microbial DNA from soil samples collected at dumping sites. The presence of distinct DNA and RNA bands highlights the effectiveness of the DNA extraction process, even in the absence of proteinase treatment. These findings provide valuable insights into the microbial diversity within dumping site soils, suggesting a rich and varied microbial community that may play significant roles in soil health and waste decomposition.

References :

  1. Fierer, N., & Jackson, R. B. (2006). "The diversity and biogeography of soil bacterial communities." Proceedings of the National Academy of Sciences, 103(3), 626-631. DOI: 10.1073/pnas.0507535103
  2. Liu, H., Carvalhais, L. C., Crawford, M., Singh, E., Dennis, P. G., Pieterse, C. M. J., & Schenk, P. M. (2017). "Inner plant values: Diversity, colonization and benefits from endophytic bacteria." Frontiers in Microbiology, 8, 2552. DOI: 10.3389/fmicb.2017.02552
  3. Wilson, K. (2001). "Preparation of genomic DNA from bacteria." Current Protocols in Molecular Biology, 56(1), 2.4.1-2.4.5. DOI: 10.1002/0471142727.mb0204s56
  4. Zhou, J., Bruns, M. A., & Tiedje, J. M. (1996). "DNA recovery from soils of diverse composition." Applied and Environmental Microbiology, 62(2), 316-322. DOI: 10.1128/aem.62.2.316-322.1996
  5. Pankhurst, C. E., Doube, B. M., Gupta, V. V. S. R., & Grace, P. R. (1997). "Soil biodiversity, bioindicators and soil health." Biodiversity and Sustainability of Soil Organisms, 15, 273-286. DOI: 10.1016/B978-0-444-82145-6.50024-4
  6. Schmidt, S. K., & Lipson, D. A. (2004). "Microbial growth under the snow: Implications for nutrient and allelochemical availability in temperate soils." Plant and Soil, 259(1-2), 1-7. DOI: 10.1023/ B.0000020931.62445.f3
  7. Strong, P. J., & Burgess, J. E. (2008). "Treatment methods for wine-related and distillery-related wastewaters: A review." Bioremediation Journal, 12(2), 70-87. DOI: 10.1080/10889860802006836

This study aims to investigate the microbial diversity present in soil samples collected from various dumping sites, which are known for their complex and heterogeneous waste environments. The primary objective is to isolate and characterize the microbial communities inhabiting these soils, providing insights into their composition and potential ecological roles. Soil samples were collected by digging 5 cm deep at multiple dumping locations to ensure a representative sampling of the microbial populations. The collected soil samples underwent a serial dilution process, extending up to 10^-9 and 10^-10 dilutions, to facilitate the isolation of individual microbial colonies. These diluted samples were then inoculated onto Standard Potato Dextrose Agar (PDA) media, a nutrient-rich medium conducive to microbial growth. Following an incubation period, distinct bacterial colonies were observed and isolated for further analysis. DNA was extracted from the isolated bacterial colonies using a standard DNA extraction protocol. The quality and purity of the extracted DNA were assessed through gel electrophoresis, a technique that separates DNA fragments based on their size. Remarkably, the electrophoretic analysis revealed clear and distinct DNA bands along with RNA bands, indicating successful extraction of high-quality microbial DNA. Notably, this was achieved without the use of proteinase treatment, which is typically employed to remove protein contaminants from DNA samples. The results of this study underscore the robustness of the methodologies employed in isolating and characterizing microbial DNA from soil samples collected at dumping sites. The presence of distinct DNA and RNA bands highlights the effectiveness of the DNA extraction process, even in the absence of proteinase treatment. These findings provide valuable insights into the microbial diversity within dumping site soils, suggesting a rich and varied microbial community that may play significant roles in soil health and waste decomposition.

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