Plant Diversity ›› 2014, Vol. 36 ›› Issue (03): 365-374.doi: 10.7677/ynzwyj201413151

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High Throughput SSR Multiplex Kits (12plex) for Euphrates’ Poplar, Populus euphratica (Salicaceae)

 XU  Fang, ZHAO  Shu, WU  Rong-Ling, DU  Fang   

  1. Center for Computational Biology, National Engineering Laboratory for Tree Breeding,
    Beijing Forestry University, Beijing 100083, China
  • Received:2013-07-10 Online:2014-05-25 Published:2013-10-12
  • Supported by:

    Doctoral Program of Higher Education of China (2011DD14120014) and Fundamental Research Funds for the Central Universities (TD 201201) to FKD

Abstract:

Multiplex PCR of microsatellite is a costeffective and highthroughput technique of genotyping. We developed a new 12plex PCR kit for Populus euphratica, the only tree species in desert area ranging from Western China to Mediterranean coast. Three primers were designed for the expressed sequence tags (ESTs) sequences from the NCBI database and the other nine primers were designed based on the EST sequences of Peuphratica obtained by Solexa. The multiplex kit was tested by 96 samples from three natural populations. The results showed sufficient amplification stability and high polymorphism. All the 12 loci used in this kit showed a high transferability (79%) in other seven species from five sections of the genus. The new 12plex kit combined with the two eight multiplex kits we had developed in previous studies, should be useful to reveal the genetic mechanism and evolution history of the Peuphratica and related species. During the research, we found that primers selection, amplification efficiency, null allele detection are the essential parts of the multiplex kit development.

Key words: Multiplex SSR, Populus euphratica, Genotyping

CLC Number: 

  • Q 78