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

Ajith Samantha Rathnayake, Josep Allué, Mercè Llugany, Anna Puig-Pujol, Kshanika Hirimburegama, Charlotte Poschenrieder

Department of Agriculture, Research and Technology Food and Agriculture Institute (IRTA)—Catalan Institute of Vine and Wine (INCAVI), Vilafranca del Penedés, Spain.
Department of Plant Sciences, Faculty of Science, University of Colombo, Colombo, Sri Lanka.
Plant Physiology Laboratory, Faculty of Biosciences, Autonomous University of Barcelona (UAB), Bellaterra, Spain.
Plant Physiology Laboratory, Faculty of Biosciences, Autonomous University of Barcelona (UAB), Bellaterra, Spain Department of Plant Sciences, Faculty of Science, University of Colombo, Colombo, Sri Lanka.

DOI: 10.4236/ajps.2014.513217                

Abstract

Use of a single seed is very useful for genetic studies on Vitis vinifera. However, molecular markers require a fair amount of high purity DNA. Grapevine contains high concentrations of polysaccharides, polyphenols, tannins and other secondary metabolites. These compounds may hamper the DNA isolation processes and subsequent analysis. In this study we have compared two DNA isolation methods: the NucleoSpin Plant II method and a modified protocol from Doyle and Doyle. The average value of 260/280 nm absorption ratio, which is used to assess the purity of DNA and RNA was 1.8 (accepted as “pure” DNA) and 0.9 (presence of protein or other contaminants) for the first and second method, respectively. Using the NucleoSpin protocol, from a single seed (20 - 35 mg) we obtained an average yield of extracted DNA of 24.8 ± 5.2 to 38.4 ± 11.5 ng·mg^-1 dry weight. Among the two protocols examined, the NucleoSpin method was more efficient and gave better quality of DNA values compared to those from the modified Doyle and Doyle procedures.

Keywords

Vitis, Single Seed, High Quality DNA, DNA Extraction Method

Conflicts of Interest

The authors declare no conflicts of interest.

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