用于MLPA技术的单链长探针制备方法研究  

A novel procedure for the preparation of the downstream long single-strand DNA probes for multiplex ligation-dependent probe amplification

作　　者：陈菲[1] 陈枝楠[2] 康林[2] 潘广[2] 叶奕优[2] 肖启明[1] 凌杏园[2] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128 [2]深圳出入境检验检疫局

出　　处：《植物检疫》2012年第2期1-5,共5页Plant Quarantine

基　　金：深圳出入境检验检疫局科技项目(SZ2008104);转基因重大专项课题(2009ZX08012-001B);深圳市科技项目(SY200609110044A)

摘　　要：依据多重连接依赖探针扩增技术(MLPA)的单链探针设计要求,将检测各基因位点的MPLA探针对的上游探针设计为短探针,采用化学法加以合成;将下游探针作为长探针,以不对称PCR方法制备。本文应用该技术制备了检测12个转基因玉米品系的MLPA下游探针;并用其中的2个长探针进行转基因玉米品系检测,结果表明制备探针完全符合转基因检测要求。该技术操作简单,成本低,应用价值高。According to the design principles of single-stranded probe in MLPA,each upstream probe of the MLPA probe pairs for different loci are designed as short probes and synthesized chemically,however,the downstream probes are designed as long oligo-nucleotides probes and prepared with asymmetric PCR.In this study,With this novel procedure,12 such probes for 12 GMO corn lines were prepared,and were proved to be effective with 2 such long probes being practically used for GMO detection.This novel procedure has the characteristic of simple manipulation and low cost,thus has high value for practical use.

关 键 词：MLPA 探针 PUC18 不对称PCR 限制性内切酶 

分 类 号：S188[农业科学—农业基础科学]