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

ELECTROPHORESIS (2016)
Jiří Drábek, Michaela Smolíková, Ruslan Kalendar, Fernando A. Lopes Pinto, Pavel Pavloušek, Karel Klepárník, Ivo Frébort
First Online: 2 October 2016
DOI: 10.1002/elps.201600068

Abstract

Although the analysis of length polymorphism at short tandem repeat (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 capillary electrophoresis with laser induced fluorescence, and tested it on 90 grape cultivars. The new protocol requires sub-nanogram quantities of the DNA template and enables automated, high-throughput genetic analysis with reasonable discriminatory power. As such, it represents a step towards further standardization of grape DNA profiling.

Keywords: Short tandem repeats; grapevine DNA analysis; multiplex PCR; Vitis vinifera L

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О нас

Республиканское государственное предприятие «Национальный центр биотехнологии» Комитета науки Министерства образования и науки Республики Казахстан – ведущий биологический научный центр страны, реализующий государственную политику поддержки и развития биотехнологической отрасли - создан Указом Президента Республики Казахстан в 1993 г.

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