基于基因组DNA混合池的EST-SSR引物快速筛选方法研究  

Studies on quick screening of EST-SSR primers based on genomic DNA mixing pool

作　　者：刘果[1] 谢耀坚[1] 张党权[2,3,4] 谷振军[2,3,4] 

机构地区：[1]国家林业局桉树研究开发中心,广东湛江524022 [2]中南林业科技大学林业生物技术湖南省重点实验室,湖南长沙410004 [3]中南林业科技大学经济林育种与栽培国家林业局重点实验室,湖南长沙410004 [4]中南林业科技大学经济林培育与保护省部共建教育部重点实验室,湖南长沙410004

出　　处：《中南林业科技大学学报》2012年第9期124-129,共6页Journal of Central South University of Forestry & Technology

基　　金：林业公益性行业专项(201104003);863课题(2011AA100202);湖南省科技厅重点项目(2010NK2008;2011NK2019);国家863计划课题(2011AA100203)

摘　　要：林木SSR标记的引物筛选工作量大,十分费时且消耗大量实验材料。采用基因组DNA混合池技术,将研究对象的若干个基因组DNA样品等量混合后,作为单一模板进行SSR的PCR扩增,能显著降低实验的工作量和成本。以选择的150对EST-SSR引物为对象,对6个桉树基因组DNA样品进行等量混合,进行PCR扩增筛选引物,结果表明,与常规筛选方法相比,基因组DNA混合池方法大幅度缩短了实验周期,显著减少了基因组DNA的消耗,可用于大量林木SSR引物的快速高效筛选。The screening of SSR primers for forestry is a heavy research which is very time consuming and consume intensive experimental materials.By employing genomic DNA pooling technology,a method of mixing several genomic DNA templates equivalently as the single template for PCR,was used to quickly screen SSR primers,which will significantly reduce the experimental workload and cost.Genomic DNA pooling constructed by equivalently mixing 6 Eucalyptus genomic DNA templates was used to screen 150 pairs of EST-SSR primers.The analytical results show that comparing with conventional screening method,the genomic DNA pooling method can greatly shorten experimental period and significantly reduce the consumption of genomic DNA,suggesting that this method is a fast and efficient method for screening substantive SSR primers used in trees.

关 键 词：基因组DNA混合池 引物筛选 EST-SSR PCR 

分 类 号：S792.39[农业科学—林木遗传育种]