Optimization of High-Yielding Protocol for Dna Extraction From-Leaves of Asam Gelugor (Garcinia Atroviridis), Malaysian High Economic Value Of Medicinal Plant

International Journal of Agriculture & Environmental Science

© 2022 by SSRG - IJAES Journal

Volume 9 Issue 3

Year of Publication : 2022

Authors : Zulkifli Ahmad Seman, Sanimah Simoh, Zaulia Othman, Nurhazwani Mustaffer, Zuraida AB Rahman, Mohd Shukri Mat Ali

10.14445/23942568/IJAES-V9I3P109

How to Cite?

Zulkifli Ahmad Seman, Sanimah Simoh, Zaulia Othman, Nurhazwani Mustaffer, Zuraida AB Rahman, Mohd Shukri Mat Ali, "Optimization of High-Yielding Protocol for Dna Extraction From-Leaves of Asam Gelugor (Garcinia Atroviridis), Malaysian High Economic Value Of Medicinal Plant," SSRG International Journal of Agriculture & Environmental Science, vol. 9,  no. 3, pp. 62-68, 2022. Crossref, https://doi.org/10.14445/23942568/IJAES-V9I3P109

Abstract:

DNA isolation is difficult in woody plants due to polysaccharides, tannins, alkaloids, polyphenols, and other secondary metabolites that interfere with isolated DNA quality. Here we report for the first time a fast, reliable, and less expensive method to isolate genomic DNA from leaves of Asam Gelugor (Garcinia atroviridis). This method generally started with a sample washing step by Triton buffer (2%) before isolating gDNA according to a modified CTAB method. We employed a high concentration 4% (w/v) CTAB, 1.5% (w/v) PVP (Polyvinylpyrrolidone), 0.3% (v/v) mercaptoethanol and relatively higher concentration of 30 mM EDTA compared to extraction method developed for other Garcinia varieties. The method has included a precipitation step by using 5M NaCl and chilled ethanol to increase the solubility of polysaccharides and 50 mM ascorbic acid to inhibit polyphenol oxidase activity. High purity of gDNA was obtained as evidenced by the A260/A280 ratio ranging from 1.7 to 1.8, which suggested that proteins and RNA did not contaminate the purified gDNA. DNA concentration ranged from 1.5 to 5.0 µg/µl. This newly modified protocol enabled the isolation of high-quality Garcinia atroviridis genomic DNA for subsequent genetic diversity study of germplasms and gender identification.

Keywords:

Garcinia Atroviridis, DNA extraction, Optimization, Leaf tissue.

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