Drábek Jiří, Smolíková Michaela, Kalendar Ruslan, Pinto Fernando A Lopes, Pavloušek Pavel, Klepárník Karel, Frébort Ivo
Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University, Olomouc, Czech Republic.
Department of Biochemistry, Palacký University, Olomouc, Czech Republic.
Electrophoresis. 2016 Dec;37(23-24):3059-3067. doi: 10.1002/elps.201600068. Epub 2016 Oct 26.
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.
尽管对STR基因座长度多态性的分析已成为葡萄品种鉴定的首选方法，但为此目的的方法标准化落后于人类和动物法医遗传学中DNA分析方法的标准化。因此，本研究的目的是设计并验证一种具有实际可用多重水平的葡萄STR检测方案。利用免费的生物信息学工具、已发表的引物序列和核苷酸数据库，我们构建并优化了一组引物，用于通过多重PCR和激光诱导荧光CE同时分析六个STR基因座（VVIi51、scu08vv、scu05vv、VVMD17、VrZAG47和VrZAG83），并在90个葡萄品种上进行了测试。新方案需要亚纳克量的DNA模板，并能够以合理的鉴别力进行自动化、高通量遗传分析。因此，它代表了葡萄DNA分析进一步标准化的一步。
