A simple and efficient protocol for high quality of DNA from Vitis quandrangularis L.

Abstract

Vitis quadrangularis L., popularly known as Hadsankal in Saurashtra region, exhibits quadrangular and flat stems structure. The present study was aimed to establish efficient protocol for DNA extraction and to evaluate taxonomic status of morphological variants using molecular characterization method RAPD. Three protocols were tested and the final protocol was modified and optimized for high quality DNA. Third protocol was CTAB and SDS based method, developed by adding higher concentration of CTAB and NaCl with an aim to remove polysaccharides, and extra PVP as a phenols and polyphenols remover. It resulted into high quality DNA with average purity of 1.8 and yield 200 g per gm stem tissue that appropriate for molecular characterization. For molecular marker analysis total 20 decamer primers of OPA series were tested and among them four primers successfully revealed 42.10 % polymorphism. Thus, the results adequately discriminate the morphological variants.