Rapid, High Quality DNA Isolation from Tunisian Grapevine (Vitis vinifera L.) Cultivars and Optimization of the RAPD Marker Technique

Abstract

Various problems are encountered during DNA extraction from Vitis species, harbouring high levels of secondary metabolites and polysaccharides. A simple and highly efficient protocol for isolating large quantities (0.5±0.3 mg.g-1 of leaf tissue) of high-quality DNA, from dry young Vitis vinifera leaves, is described in the present study. Thus, three different DNA extraction protocols were examined. The isolated DNA is essentially free of polysaccharides, polyphenols, and other major contaminants as judged by viscosity, clear color, A260/280 ratio, and RAPD suitability. Moreover, the RAPD profiling from the isolated DNA was optimized to produce scorable and clear amplicons in all studied cultivars