Original Research
High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas
Submitted: 26 September 2024 | Published: 23 January 2025
About the author(s)
Shiwani Singh, Department of Biotechnology, Manav Rachna Centre for Medicinal Plant Pathology, Manav Rachna International Institute of Research and Studies, Faridabad, Haryana, IndiaManeesh S. Bhandari, Genetics & Tree Improvement Division, ICFRE-Forest Research Institute, Dehradun, Uttarakhand, India
Neeraj Dwivedi, Department of Biotechnology, Manav Rachna Centre for Medicinal Plant Pathology, Manav Rachna International Institute of Research and Studies, Faridabad, Haryana, India
Abstract
Background: Desmodium gangeticum, a crucial medicinal shrub from the Himalayan Shivalik region, is an economically important species globally. It contains a vast range of secondary metabolites, majorly polyphenols, that interfere with high-quality deoxyribonucleic acid (DNA) extraction by binding their oxidised form to DNA covalently and rendering it unusable for molecular research.
Aim: This study was focused on developing a protocol for the isolation of high-quality DNA from D. gangeticum, which is essential for carrying out molecular biological experiments. Also, the isolation of DNA in purified form has its significance in facilitating precise genetic analysis such as sequencing, restriction digestion and polymerase chain reaction (PCR)-based trials.
Setting: This study was conducted under controlled laboratory environment at Manav Rachna Centre for Medicinal Plant Pathology, Department of Biotechnology, Manav Rachna International Institute of Research and Studies, Faridabad, India.
Methods: Advanced techniques, commercial kits and rigorous modifications in existing DNA extraction buffer conditions and extraction protocols from both fresh and dried leaves are considered to optimise high-quality DNA isolation.
Results: With the standardised buffer conditions and protocol, high-quality DNA with purity indicated by a 260/280 nm absorbance ratio in a range of 1.8 to 2.0 with 250 ng/µL – 950 ng/µL concentration from 0.5 g of leaf sample and also validated by agarose gel electrophoresis showing reduced contaminations and intact DNA was isolated.
Conclusion: This standardised buffer condition and protocol led to high-quality DNA isolation, which was found good for restriction digestion using endonuclease enzymes, PCR and other molecular biology techniques, highlighting the ongoing efforts to enhance the reliability of genetic studies in medicinal plants.
Contribution: This study adds to the existing knowledge about the strategies and procedures for extracting high-quality DNA and good yield from leaf samples of D. gangeticum.
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