Isolation and Purification of Genomic DNA from Black Plum (Eugenia Jambolana Lam.) for Analytical Applications

Medicinal plants and tree species contains high amount of secondary metabolites in their tissues, which acts as contaminants interfering analytical quality DNA isolation. Most of the efforts end up with colored or viscous preparations. An attempt was made to localize these contaminants in petiole tissues to emphasize deveining as important pre-processing step in medicinal plant DNA isolation procedure. Seven methods based on CTAB, SDS and Triton –X-100 combined with PVP and β-mercaptoethanol was used to identify suitable method for genomic DNA isolation from Black Plum (Eugenia jambolana Lam.) leaves. Fixing leaf sample in absolute alcohol, grinding in liquid nitrogen and higher concentration of PVP (MW 40,000) i.e. 2% and β-mercaptoethanol (2%) was found to be suitable. The DNA quality was further enhanced by purification using DEAE cellulose column chromatography. It was observed that, this modified CTAB method in combination with chromatographic purification yields DNA with sufficient quantity and quality for analytical applications like PCR amplification and cloning. CTAB

Cetyl_trimethylammonium_bromide

Polyvinylp-pyrrolidone

Sodium dodecyl sulfate

Di Ethyl Amino Ethyl cellulose