干旱区生态环境知识资源中心(Arid): 一种优化的胡杨高效多重(12重)SSR体系

Arid

	一种优化的胡杨高效多重(12重)SSR体系
其他题名	High Throughput SSR Multiplex Kits (12-plex) for Euphrates’ Poplar, Populus euphratica (Salicaceae)
	徐放; 赵舒; 邬荣领; 杜芳
来源期刊	植物分类与资源学报
ISSN	2095-0845
出版年	2014
卷号	36 期号:3 页码:365-374
中文摘要	微卫星多重PCR方法是一种非常经济并且高通量的基因分型技术。本研究在耐干旱、盐碱的胡杨(Populus euphratica)中开发出一套荧光标记的12重微卫星工作体系。该体系包含12条表达序列标签微卫星(EST-SSR)引物,其中3条设计于NCBI,另外9条设计于二代的转录组序列。利用该多重微卫星体系可在单一的PCR反应体系中成功扩增出12条表达序列标签的微卫星短序列片段,并在胡杨的3个自然居群96个个体中对该体系进行了验证,结果显示该体系具有很高的稳定性及多样性。同时,在杨属的5个派7个种中对其通用性进行了检验,显示这些引物具有很高的通用性,成功扩增率为79%。本研究中提供的12重多重PCR结合本实验已经公开发表的2个8重体系对揭示胡杨及其他杨树的进化历史具有重要的作用。最后,本研究认为引物的选择,扩增效率,哑等位基因的检测是多重体系开发过程中最为关键的步骤。
英文摘要	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.
中文关键词	胡杨 ; 多重PCR ; 基因分型
英文关键词	Multiplex SSR Populus euphratica Genotyping
语种	英语
国家	中国
收录类别	CSCD
WOS类目	GENETICS HEREDITY
WOS研究方向	Genetics & Heredity
CSCD记录号	CSCD:5153581
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/232009
作者单位	Center for Computational Biology, Beijing Forestry University, National Engineering Laboratory for Tree Breeding, Beijing 100083, China
推荐引用方式 GB/T 7714	徐放,赵舒,邬荣领,等. 一种优化的胡杨高效多重(12重)SSR体系[J],2014,36(3):365-374.
APA	徐放,赵舒,邬荣领,&杜芳.(2014).一种优化的胡杨高效多重(12重)SSR体系.植物分类与资源学报,36(3),365-374.
MLA	徐放,et al."一种优化的胡杨高效多重(12重)SSR体系".植物分类与资源学报 36.3(2014):365-374.