High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas
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 th...
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| Format: | Article |
| Language: | English |
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AOSIS
2025-01-01
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| Series: | Journal of Medicinal Plants for Economic Development |
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| Online Access: | https://jomped.org/index.php/jomped/article/view/272 |
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| author | Shiwani Singh Maneesh S. Bhandari Neeraj Dwivedi |
| author_facet | Shiwani Singh Maneesh S. Bhandari Neeraj Dwivedi |
| author_sort | Shiwani Singh |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-a2d699980d144d319efa522ae1a49d6f |
| institution | Kabale University |
| issn | 2519-559X 2616-4809 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | AOSIS |
| record_format | Article |
| series | Journal of Medicinal Plants for Economic Development |
| spelling | doaj-art-a2d699980d144d319efa522ae1a49d6f2025-02-11T13:24:35ZengAOSISJournal of Medicinal Plants for Economic Development2519-559X2616-48092025-01-0191e1e710.4102/jomped.v9i1.272120High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik HimalayasShiwani Singh0Maneesh S. Bhandari1Neeraj Dwivedi2Department of Biotechnology, Manav Rachna Centre for Medicinal Plant Pathology, Manav Rachna International Institute of Research and Studies, Faridabad, HaryanaGenetics & Tree Improvement Division, ICFRE-Forest Research Institute, Dehradun, UttarakhandDepartment of Biotechnology, Manav Rachna Centre for Medicinal Plant Pathology, Manav Rachna International Institute of Research and Studies, Faridabad, HaryanaBackground: 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.https://jomped.org/index.php/jomped/article/view/272desmodium gangeticumdna isolationmedicinal plantrestriction digestionmodified ctabsecondary metabolitepolyphenolspvpp |
| spellingShingle | Shiwani Singh Maneesh S. Bhandari Neeraj Dwivedi High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas Journal of Medicinal Plants for Economic Development desmodium gangeticum dna isolation medicinal plant restriction digestion modified ctab secondary metabolite polyphenols pvpp |
| title | High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas |
| title_full | High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas |
| title_fullStr | High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas |
| title_full_unstemmed | High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas |
| title_short | High-quality DNA extraction for Desmodium gangeticum: A medicinal shrub of Shivalik Himalayas |
| title_sort | high quality dna extraction for desmodium gangeticum a medicinal shrub of shivalik himalayas |
| topic | desmodium gangeticum dna isolation medicinal plant restriction digestion modified ctab secondary metabolite polyphenols pvpp |
| url | https://jomped.org/index.php/jomped/article/view/272 |
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