| 转基因油菜RT73品系特异性检测标准分子协同实验验证 |
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| 引用本文： | 李想,褚庆华,高琴,吕蓉,潘良文,李俊毅,张其刚. 转基因油菜RT73品系特异性检测标准分子协同实验验证[J]. 中国农业科技导报, 2011, 13(6): 33-40. DOI: 10.3969/j.issn.1008-0864.2011.06.06 |
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| 作者姓名： | 李想 褚庆华 高琴 吕蓉 潘良文 李俊毅 张其刚 |
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| 作者单位： | (1.上海出入境检验检疫局, 上海 200135,2.华东理工大学, 上海 200237) |
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| 基金项目： | 国家科技支撑计划项目(2008BAK41B01-2); 国家转基因生物新品种培育重大专项(2008ZX08012-001;2009ZX08012-002B); 上海市科委研发平台专项(10DZ2294102)资助 |
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| 摘 要： | 对构建的适于转基因油菜RT73品系特异性检测标准分子pEASY-RT73在普通PCR和实时荧光PCR检测中的适用性进行了实验室间协同实验验证。对8家实验室结果的统计分析表明,标准分子pEASY-RT73对转基因油菜RT73品系特异性检测及油菜内源基因HMG和PEP检测具有高特异性。在普通PCR扩增中,标准分子对PEP和RT73品系特异性序列的检测下限均为20拷贝,对HMG基因的检测下限为50拷贝;在实时荧光PCR扩增中,HMG和RT73品系特异性序列的检测下限均为10拷贝,PEP检测下限为50拷贝。虽然各实验室所用仪器和试剂有较大差异,但对检测结果影响不大。因此,标准分子pEASY-RT73可稳定的作为新型标准物质用于转基因油菜RT73品系特异性普通PCR和实时荧光PCR检测。
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| 关 键 词： | RT73 标准分子 品系特异性 普通PCR 实时荧光PCR, |
Inter-laboratory Validation of a Plasmid Reference Molecule Suitable for Genetically Modified Canola RT73 Event-specific Detection |
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| LI Xiang , ZHU Qing-hua , GAO Qin , LV Rong , PAN Liang-wen , LI Jun-yi , ZHANG Qi-gang. Inter-laboratory Validation of a Plasmid Reference Molecule Suitable for Genetically Modified Canola RT73 Event-specific Detection[J]. Journal of Agricultural Science and Technology, 2011, 13(6): 33-40. DOI: 10.3969/j.issn.1008-0864.2011.06.06 |
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| Authors: | LI Xiang ZHU Qing-hua GAO Qin LV Rong PAN Liang-wen LI Jun-yi ZHANG Qi-gang |
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| Affiliation: | (1.Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135|2.East China University of Science and Technology, Shanghai, 200237, China) |
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| Abstract: | In this study,the applicability of a reference molecule pEASY-RT73 suitable for event-specific detection of genetically modified(GM) canola RT73 in conventional and real-time PCR assays was validated by inter-laboratory trail.The analysis of 8 labs reports showed that the plasmid pEASY-RT73 was highly specific to GM canola RT73 rapeseed endogenous gene HMG and PEP detections.In qualitative PCR,the limits of detection(LODs) that could be detected by all 8 participants were 20 copies of plasmid DNA for PEP an... |
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| Keywords: | RT73 reference molecule event-specific conventional PCR real-time PCR |
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