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Direct multiplex PCR for grapevine genotyping and varietal identification  

2013-02-03 15:45:36|  分类: 数量遗传学 |  标签: |举报 |字号 订阅

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Abstract

Grapevine cultivar identification is based mainly on two complementary methodologies: microsatellite (simple sequence repeat (SSR)) DNA analysis and traditional ampelography. Here, we report a direct multiplex PCR approach that allows the simultaneous amplification of 11 SSR loci from crude samples, i.e. bypassing DNA extraction, by using an engineered DNA polymerase improved to tolerate plant PCR inhibitors. Many different plant tissues were successfully amplified: leaf, root, wood, berry flesh and skin, stalk and must, from wine and table grape varieties, and rootstocks. The direct multiplex PCR that we propose is quicker and cheaper than the methodologies used until now, and provides accurate results. Thus, SSR DNA analysis becomes economically more accessible to a larger number of potential users in addition to research institutes.

Direct multiplex PCR for grapevine genotyping and varietal identification - 喜欢吃桃子 - wangyufeng的博客
 

Fig. 1. Example of the multiplex PCR molecular profile of 11 SSR loci obtained with the proposed methodology. (a) NED-,
(b) 6-FAM-, (c) PET- and (d) VIC-labelled SSR loci. Grey and red stripes represent the home-made bin set produced with
reference varieties.


Full text:http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8771889

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