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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,
Jazyk angličtina Země Německo
Typ dokumentu časopisecké články
PubMed
27696463
DOI
10.1002/elps.201600068
Knihovny.cz E-zdroje
- MeSH
- algoritmy MeSH
- DNA rostlinná analýza genetika MeSH
- genetické markery genetika MeSH
- mikrosatelitní repetice genetika MeSH
- multiplexová polymerázová řetězová reakce metody MeSH
- reprodukovatelnost výsledků MeSH
- víno MeSH
- Vitis klasifikace genetika MeSH
- výpočetní biologie MeSH
- Publikační typ
- časopisecké články 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
Citace poskytuje Crossref.org
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