TITLE:
High Quality DNA Obtained from a Single Seed of Vitis vinifera L. Using Rapid DNA Extraction Method
AUTHORS:
Ajith Samantha Rathnayake, Josep Allué, Mercè Llugany, Anna Puig-Pujol, Kshanika Hirimburegama, Charlotte Poschenrieder
KEYWORDS:
Vitis, Single Seed, High Quality DNA, DNA Extraction Method
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.5 No.13,
June
24,
2014
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.