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. Thepresent study was aimed to establish efficient protocol for DNA extraction and to evaluate taxonomic status of morphologicalvariants using molecular characterization method RAPD. Three protocols were tested and the final protocol was modified andoptimized for high quality DNA. Third protocol was CTAB and SDS based method, developed by adding higher concentration ofCTAB and NaCl with an aim to remove polysaccharides, and extra PVP as a phenols and polyphenols remover. It resulted intohigh 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 successfullyrevealed 42.10 % polymorphism. Thus, the results adequately discriminate the morphological variants.

Keywords

• Vitis quadrangularis L.
• DNA extraction
• Molecular Characterization
• RAPD