Extraction of High-quality Genomic DNA Using Species Tailored Protocol for Prosopis cineraria (L.) Druce (Khejri)

Abstract

To effectively conserve genetic diversity in forestry species, molecular methods are essential. These methods necessitate high-quality genomic DNA as a starting material which contains the complete genetic information of an organism, and by extracting it, scientists can access and analyze this information. This information is essential for developing effective conservation strategies and understanding the complex processes of evolution. The objective of this study was to develop a protocol for extracting High-Quality genomic DNA from the arid tree species Prosopis cineraria. The current extraction protocol is based upon the conventional cetyl trimethyl ammonium bromide (CTAB) method with further modifications for the extraction of DNA. The principle modifications currently employed for DNA extraction involved the use of higher concentration of CTAB and sodium chloride. Additionally, L-ascorbic acid and Polyvinylpyrrolidone (PVP) was also added. The incubation time was doubled and DNA was precepted in 100% ice-cold ethanol. Further a RNAse treatment was also included for 30 mins. The yield of DNA ranged from 630-4200ng/µl with average yield of 2041ng/µl, and the absorbance lied between 1.6-2.15 for A260/280 and 1.64-2.31 for A260/230, indicating minimal levels of contaminating metabolites. This method offers a simple, efficient, and cost-effective way to extract high-quality genomic DNA, making it ideal for large-scale genetic studies.