Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars

0465488 - ÚIACH 2017 RIV DE eng J - Journal Article
Drábek, A. - Smolíková, M. - Kalendar, R. - Pinto, F. A. L. - Pavloušek, P. - Klepárník, Karel - Frébort, I.
Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars.
Electrophoresis. Roč. 37, 23-24 (2016), s. 3059-3067. ISSN 0173-0835. E-ISSN 1522-2683
Institutional support: RVO:68081715
Keywords : grapevine DNA analysis * multiplex PCR * STRs * Vitis vinifera L
Subject RIV: CB - Analytical Chemistry, Separation
Impact factor: 2.744, year: 2016

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.
Permanent Link: http://hdl.handle.net/11104/0264037