An effective and low-cost method for DNA extraction from Rheum species (A medicinal herb) without using liquid nitrogen for RAPD/ISSR Study

Abstract

Rheum is an important medicinal plant belonging to family Polygonaceae and is found to have anti-microbial, anti-purgative and anti-cancer properties. The objective of the present study is to standardize an effective and low-cost method for the extraction of DNA from the young juvenile leaves of Rheum plant obtained from the Uttarakhand region. The protocol involved the extraction of DNA using a modified CTAB method without the use of any liquid nitrogen. The concentration of NaCl used in extraction buffer was slightly higher (1.4M) to remove polysaccharides and varying ratios of Beta-mercaptoethanol used to remove the secondary metabolites and keep them away from interfering the extraction process. The absorbance ratio of the extracted DNA at A260/280 nm was within the permissible range of 1.7-2.0, indicating the purity of DNA and showing that the DNA is contamination free. Its concentration was approximately 1485 ng/μl and it was found suitable for molecular-based study.