A Fast, Efficient Method for Extracting DNA from Leaves, Stems, and Seeds of Vitis vinifera L.

Abstract

A new method to extract DNA that can be applied to the leaves, seeds, and stems of Vitis vinifera uses a combination of dodecyltrimethylammonium bromide (DTAB) and cetyltrimethylammonium bromide (CTAB) and allows for optimal DNA extraction from recalcitrant tissues. The technique is as rapid as commercial kits and requires only small amounts of material. As it does not require RNase treatment, its application is approximately one-eighth of the cost. The possibility of applying the same DNA extraction method to different tissues is advantageous for two reasons: it diminishes a source of heterogeneity and it can be applied to samples obtained during any season of the year, including winter. The proposed method provides high-quality DNA with a good yield from the leaves, seeds, and stems of V. vinifera and guarantees identical results of posterior SSR analyses, regardless of the tissue used and the stage of ripening at which it was collected. Furthermore, since this method can be applied to stems, it can be used as a first step for quality control in grapevine nurseries.