| 用于MLPA技术的单链长探针制备方法研究 |
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| 引用本文： | 陈菲,陈枝楠,康林,潘广,叶奕优,肖启明,凌杏园.用于MLPA技术的单链长探针制备方法研究[J].植物检疫,2012(2):1-5. |
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| 作者姓名： | 陈菲 陈枝楠 康林 潘广 叶奕优 肖启明 凌杏园 |
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| 作者单位： | 湖南农业大学生物科学技术学院;深圳出入境检验检疫局 |
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| 基金项目： | 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) |
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| 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。
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| 关 键 词： | MLPA 探针 pUC18 不对称PCR 限制性内切酶 |
A novel procedure for the preparation of the downstream long single-strand DNA probes for multiplex ligation-dependent probe amplification |
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| Chen Fei,Chen Zhinan,Kang Lin,Pan Guang,Ye Yiyou,Xiao Qiming,Ling Xingyuan.A novel procedure for the preparation of the downstream long single-strand DNA probes for multiplex ligation-dependent probe amplification[J].Plant Quarantine,2012(2):1-5. |
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| Authors: | Chen Fei Chen Zhinan Kang Lin Pan Guang Ye Yiyou Xiao Qiming Ling Xingyuan |
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| Institution: | 1.Life Science and Technology College,Hunan Agriculture University,Changsha 410128,china;2.Shenzhen Entry-Exit Inspection and Quarantine Bureau) |
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| Abstract: | 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. |
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| Keywords: | MLPA(multiplex ligation-dependent probe amplification) probe pUC18 asymmetric PCR restriction enzyme |
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