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Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars
J. Drábek, M. Smolíková, R. Kalendar, FA. Pinto, P. Pavloušek, K. Klepárník, I. Frébort,
Language English Country Germany
Document type Journal Article
- MeSH
- Algorithms MeSH
- DNA, Plant analysis genetics MeSH
- Genetic Markers genetics MeSH
- Microsatellite Repeats genetics MeSH
- Multiplex Polymerase Chain Reaction methods MeSH
- Reproducibility of Results MeSH
- Wine MeSH
- Vitis classification genetics MeSH
- Computational Biology MeSH
- Publication type
- Journal Article MeSH
Although the analysis of length polymorphism at STR loci has become a method of choice for grape cultivar identification, the standardization of methods for this purpose lags behind that of methods for DNA profiling in human and animal forensic genetics. The aim of this study was thus to design and validate a grapevine STR protocol with a practically useful level of multiplexing. Using free bioinformatics tools, published primer sequences, and nucleotide databases, we constructed and optimized a primer set for the simultaneous analysis of six STR loci (VVIi51, scu08vv, scu05vv, VVMD17, VrZAG47, and VrZAG83) by multiplex PCR and CE with laser-induced fluorescence, and tested it on 90 grape cultivars. The new protocol requires subnanogram quantities of the DNA template and enables automated, high-throughput genetic analysis with reasonable discriminatory power. As such, it represents a step toward further standardization of grape DNA profiling.
Department of Biochemistry Palacký University Olomouc Czech Republic
Department of Photochemistry and Molecular Science Uppsala University Uppsala Sweden
Department of Viticulture and Enology Mendel University Brno Czech Republic
Institute of Analytical Chemistry of the ASCR Brno Czech Republic
References provided by Crossref.org
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