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实时荧光定量PCR技术在球毛壳ND35定殖检测中的应用
引用本文:刘璇,戴杨,高克祥,米士伟,吴海燕,何邦令. 实时荧光定量PCR技术在球毛壳ND35定殖检测中的应用[J]. 山东农业科学, 2012, 44(6): 17-19
作者姓名:刘璇  戴杨  高克祥  米士伟  吴海燕  何邦令
作者单位:山东农业大学植物保护学院,山东泰安,271018
基金项目:国家自然科学基金,公益性行业(农业)科研专项,山东省科技发展计划项目
摘    要:对球毛壳(Chaetomium globosum)ND35菌株的RAPD扩增特异性DNA片段进行分析,利用Prim-er 5.0软件设计了一对引物,即ND35-1:5'-GCTCGCAGCTCTGGTGCGGT-3',ND35-2:5'-GCCGATTGC-CCATGTCCACCTC-3',以球毛壳ND35菌株基因组DNA为模板,利用此引物建立并优化了SYBR GreenⅠ实时荧光定量PCR反应的最佳体系,并据此对杨树和黄瓜两种植物中ND35菌株的定殖情况进行了检测。结果表明,该引物可以用于球毛壳ND35的实时荧光定量PCR检测。

关 键 词:球毛壳ND35  实时荧光定量PCR  定殖  检测

Application of Quantitative Real - Time PCR in Colonization Detection of Chaetomium globosum ND35
LIU Xuan , DAI Yang , GAO Ke-xiang , MI Shi-wei , WU Hai-yan , HE Bang-ling. Application of Quantitative Real - Time PCR in Colonization Detection of Chaetomium globosum ND35[J]. Shandong Agricultural Sciences, 2012, 44(6): 17-19
Authors:LIU Xuan    DAI Yang    GAO Ke-xiang    MI Shi-wei    WU Hai-yan    HE Bang-ling
Affiliation:(College of Plant Protection,Shandong Agricultural University,Taian 271018,China)
Abstract:Based on the analysis of RAPD specific DNA fragment of Chaetomium globosum ND35,one pair of primers(ND35-1:5′-GCTCGCAGCTCTGGTGCGGT-3′,ND35-2: 5′-GCCGATTGCCCATGTCCACCTC-3′) was designed by software Primer 5.0.Taking the genomic DNA of C.globosum ND35 as template,a system of quantitative real-time PCR with SYBR Green Ⅰ was developed and optimized,and then it was used to detect the colonization of strain ND35 in cucumber and poplar.The results showed that this pair of specific primers could be used for the determination of C.globosum ND35.
Keywords:Chaetomium globosum ND35  Quantitative real-time PCR  Colonization  Detection
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