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Design and validation of an STR hexaplex assay for DNA profiling of grapevine cultivars
Palacky Univ, Fac Med & Dent, Inst Mol & Translat Med, Hnevotinska 5, Olomouc 77900, Czech Republic..
Palacky Univ, Dept Biochem, Olomouc, Czech Republic..
Univ Helsinki, LUKE BI Plant Genome Dynam, Inst Biotechnol, Helsinki, Finland.;Minist Educ & Sci Republ Kazakhstan, Sci Comm, RSE Natl Ctr Biotechnol, Astana, Kazakhstan..ORCID iD: 0000-0003-3986-2460
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Photochemistry and Molecular Science.
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2016 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 37, no 23-24, 3059-3067 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2016. Vol. 37, no 23-24, 3059-3067 p.
Keyword [en]
Grapevine DNA analysis, Multiplex PCR, STRs, Vitis vinifera L
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-316142DOI: 10.1002/elps.201600068ISI: 000392421400003PubMedID: 27696463OAI: oai:DiVA.org:uu-316142DiVA: diva2:1078344
Available from: 2017-03-03 Created: 2017-03-03 Last updated: 2017-11-29Bibliographically approved

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Lopes Pinto, Fernando A.

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