An efficient DNA isolation protocol for Cymbopogon species suitable for diverse downstream applications

Abstract: Extraction of DNA from medicinal and aromatic plants is often problematic, since these plants contain high levels of secondary metabolites which interfere with PCR based downstream applications and restriction digestions. Removal of these secondary metabolites requires appropriate reagents for DNA isolation. This investigation optimised an efficient DNA isolation protocol for Cymbopogon species that yielded sufficient quantity of DNA and could be used for diverse molecular applications. The modified protocol was also compared with the two existing DNA extraction procedures for cost effectiveness, time efficiency and quality DNA recovery. The modified protocol yields good amount of DNA ranging from 76 to 90 µg/ g of fresh tissues which was significantly higher in comparison to the other two protocols. A260/A280 ratio of the DNA obtained from the modified method ranged from 1.81 to 1.87 indicates purity of DNA and was also found to be suited for downstream applications such as restriction digestions. Subsequent RAPD, ISSR, SSR and barcode gene amplification analysis suggested that the DNA isolated by our modified method was suitable for various molecular research applications. The efficiency of this method in terms of lesser time requirement and cost effectiveness makes the present method a noticeable alternative for total cellular DNA extraction for Cymbopogon species and could be adoptable by the developing countries across the world.