Abstract
High-quality, humic-acid-free pure DNA is a prerequisite for functional and sequence-based approaches of metagenomics. In the present investigation, an improved extraction buffer was developed by making a combination of powdered activated charcoal (2%; w/v), polyvinyl poly pyrrolidone (2%; w/v), and CaCl2 (2%; w/v). This trio significantly improved the purity and yield of the metagenomic DNA from the hot spring’s hot and alkaline soil. The quality of extracted metagenomic DNA was successfully validated by PCR amplification and restriction enzymes. Besides, the thermophilic amylase encoding genes were also retrieved from these soil DNA samples. Extreme habitats I harbour low microbial biomass and, therefore, demand in-situ lysis of the microbial cells to access their genomes. The protocol can potentially extract DNA from geothermal spring habitats where the count of microbial cells is low.
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Acknowledgements
The UGC-Senior Research Fellowship given to Ms Garima Chauhan, UGC-non-NET PhD Fellowship given to Ms Meghna Arya from BBAU and Project Assistant Fellowship from CST-UP to Mr Vikas Kumar are duly acknowledged. The funding for this research work was given to Dr Monica Sharma by the Uttar Pradesh Council of Science & Technology (Grant number CST/8275).
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Chauhan, G., Arya, M., Kumar, V. et al. An improved protocol for metagenomic DNA isolation from low microbial biomass alkaline hot-spring sediments and soil samples. 3 Biotech 14, 34 (2024). https://doi.org/10.1007/s13205-023-03824-4
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DOI: https://doi.org/10.1007/s13205-023-03824-4