title: Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars.
abstract: 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.
authors.affiliation: Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University, Olomouc, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Drábek
authors.affiliation: Department of Biochemistry, Palacký University, Olomouc, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Smolíková
authors.affiliation: Institute of Biotechnology, LUKE/BI Plant Genome Dynamics, University of Helsinki, Helsinki, Finland.|RSE "National Center for Biotechnology" under the Science Committee, Ministry of Education and Science of the Republic of Kazakhstan, Astana, Kazakhstan.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Kalendar
authors.affiliation: Department of Photochemistry and Molecular Science, Uppsala University, Uppsala, Sweden.
authors.forename: Fernando A Lopes
authors.initials: FA
authors.lastname: Pinto
authors.affiliation: Department of Viticulture and Enology, Mendel University, Brno, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Pavloušek
authors.affiliation: Institute of Analytical Chemistry of the ASCR, Brno, Czech Republic.
authors.forename: Karel
authors.initials: K
authors.lastname: Klepárník
authors.affiliation: Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors.forename: Ivo
authors.initials: I
authors.lastname: Frébort
mesh_terms: D000465:Algorithms; D019295:Computational Biology; D018744:DNA, Plant / Q000032:analysis* / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics; D018895:Microsatellite Repeats / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D027843:Vitis / Q000145:classification / Q000235:genetics*; D014920:Wine
chemical_list: D018744:DNA, Plant; D005819:Genetic Markers
keywords: Grapevine DNA analysis; Multiplex PCR; STRs; Vitis vinifera L