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Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars
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SYSNO ASEP 0465488 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars Author(s) Drábek, A. (CZ)
Smolíková, M. (CZ)
Kalendar, R. (FI)
Pinto, F. A. L. (SE)
Pavloušek, P. (CZ)
Klepárník, Karel (UIACH-O) RID, ORCID
Frébort, I. (CZ)Number of authors 7 Source Title Electrophoresis. - : Wiley - ISSN 0173-0835
Roč. 37, 23-24 (2016), s. 3059-3067Number of pages 9 s. Publication form Print - P Language eng - English Country DE - Germany Keywords grapevine DNA analysis ; multiplex PCR ; STRs ; Vitis vinifera L Subject RIV CB - Analytical Chemistry, Separation Institutional support UIACH-O - RVO:68081715 UT WOS 000392421400003 EID SCOPUS 84995791191 DOI 10.1002/elps.201600068 Annotation Although the analysis of length polymorphism at STR loci has become amethod 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. Workplace Institute of Analytical Chemistry Contact Iveta Drobníková, drobnikova@iach.cz, Tel.: 532 290 234 Year of Publishing 2017
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