Indian Journal of Ecology

In the present study, leaves of Sal were collected from natural populations to isolate the genomic DNA. Unfortunately, genomic DNA extracted using protocols developed byAgbagwa et al (2012) and Doyle and Doyle (1987) was inadequate and failed to obtain genomic DNAof high quality and quantity. Woody tree species are rich in higher concentrations of polyphenols, polysaccharides, and secondary metabolites and interfere in the genomic DNA isolation process. To obtain the genomic DNA in good quality and quantity, the cetyl-trimethyl ammonium bromide (CTAB) method was modified. Modifications were carried out in the concentration of PVP and 2-mercaptoethanol and the process of utilising these reagents. The quantity of extracted DNA was evaluated through a NanoBio spectrophotometer (Analytical Technologies Ltd.), and the quality was checked by 0.8% agarose gel electrophoresis. The standardised protocol yielded high molecular weight DNAin the range of 830.12 ng µl^-1 to 1597. 23 ng µl^-1 with an average of 1278.28 ng µl^-1. The absorbance ratio at 260 to 280 nm ranged from 1.81 to 1.90, with an average of 1.84, which confirmed the purity of isolated DNAand indicated the presence of very low levels of protein, RNA, and polysaccharide contaminants. The extracted DNA was used to optimise the PCR conditions for microsatellite markers and obtain sharp polymorphic bands in an agarose gel.