| 浙贝母特异性PCR鉴定方法研究 |
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| 引用本文： | 李敏,黄龙妹,赵欣,陈强.浙贝母特异性PCR鉴定方法研究[J].中草药,2014,45(12):1754-1757. |
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| 作者姓名： | 李敏 黄龙妹 赵欣 陈强 |
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| 作者单位： | 浙江工业大学药学院, 浙江 杭州 310032;浙江工业大学药学院, 浙江 杭州 310032;浙江工业大学药学院, 浙江 杭州 310032;浙江工业大学药学院, 浙江 杭州 310032 |
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| 摘 要： | 目的利用特异性引物对浙贝母Fritillaria thunbergii进行特异性PCR鉴定。方法回收RAPD扩增筛选到的浙贝母分子标记ZB1基因,将其连接进入T-载体克隆并测序。根据测序结果设计一对特异性引物P2/P3,以贝母基因组DNA为模板进行特异性PCR扩增。结果特异性PCR鉴定结果为浙贝母在约750 bp处出现特异性条带,其他贝母品种则未出现条带。结论浙贝母特异性PCR鉴定方法操作简单,准确、灵敏、重复性好,具有较好的应用前景。
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| 关 键 词： | 浙贝母 RAPD 分子鉴定 特异性PCR 分子标记 |
| 收稿时间： | 2013/12/7 0:00:00 |
Specific PCR identification of Fritillaria thunbergii |
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| LI Min,HUANG Long-mei,ZHAO Xin and CHEN Qiang.Specific PCR identification of Fritillaria thunbergii[J].Chinese Traditional and Herbal Drugs,2014,45(12):1754-1757. |
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| Authors: | LI Min HUANG Long-mei ZHAO Xin and CHEN Qiang |
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| Institution: | College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China |
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| Abstract: | Objective To identify Fritillaria thunbergii using special primer by specific PCR. Methods Gene footprint of F. thunbergii named ZB1 was screened from RAPD amplification. Reclaimed ZB1 gene was inserted into T-vector to be cloned and sequenced. One pair of specific primers P2/P3 were designed according to the ZB1 sequence, and applied in specific PCR reaction using genome DNA of F. thunbergii as template. Results A specific band around 750 bp was detected in F. thunbergii, while nothing appeared in other varieties. Conclusion The method is convenient, reproducible, and precise, with broad application prospects. |
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| Keywords: | Fritillaria thunbergii Miq RAPD molecular identification specific PCR molecular marker |
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