Comparison between modified DNA extraction protocols and commercial isolation kits in grapevine (Vitis vinifera L.)

Abstract

ABSTRACT. Various protocols have been developed and used for DNA extraction in grapevine. However, owing to the long duration of the isolation steps in previously developed protocols, researchers have preferred to use isolation kits for studies in recent years. In our study, the DNA yield and purity obtained using six methods -namely three DNA isolation protocols and three commercial DNA isolation kits -were compared. Modifications were made and the isolation steps were shortened in the previously developed DNA isolation protocols to achieve more rapid and practical protocols. The samples were taken from plants grown under vineyard and greenhouse conditions in two periods during spring and autumn. The best results among the six DNA isolation methods were discussed. The results were also supported with polymerase chain reaction analyses conducted with isolated DNAs.

Research Article An optimized protocol for DNA extraction in plants with a high content of secondary metabolites, based on leaves of Mimosa tenuiflora (Willd.) Poir. (Leguminosae).

“…To provide a reliable molecular analysis in plants, the conditions of DNA extraction should be tested once a standard protocol applicable to all plant species remains unavailable (Tacuatiá et al, 2012;Anuradha et al, 2013). Therefore, studies of genetic diversity, population structure, phylogenetics, and inheritance patterns of plants usually start from developing a suitable method of DNA extraction for a species or group of species (Akkurt, 2012;Li et al, 2013).…”

High Quality DNA Obtained from a Single Seed of <i>Vitis vinifera</i> L. Using Rapid DNA Extraction Method

“…There are many recent papers focusing on the improvement of DNA extraction methods to increase yield and purity as well as to reduce experimental time but they still use a fair amount of sample material: 0.05 -0.1 g young leaf [18] [19] or 0.2 g seeds [20]. To our best knowledge, there is only one report on DNA extraction from single seeds of Vitis vinifera [21].…”

SILEX: A fast and inexpensive high-quality DNA extraction method suitable for multiple sequencing platforms and recalcitrant plant species

“…However, commercial kits are usually expensive, with reagent costs commonly ranging between 2 and 9 US$ per sample [8,28], and many times provide low yields, insu cient for some NGS applications [8,29]. Furthermore, for some commercial kits, the DNA quality and quantity obtained in recalcitrant species is low [30][31][32]. DNA extraction methods relying on silica matrices and chaotropic salts have been reported [33][34][35][36]; however, chaotropic salts can inhibit subsequent enzymatic reactions which are essential for NGS applications [37][38][39].…”