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多重等位基因特异性PCR在人参和西洋参鉴别中的应用

[Application of multiplex allele-specific PCR for authentication of Panax ginseng and P. quinquefolius].

作者信息

Cui Guang-hong, Tang Xiao-jing, Huang Lu-qi

机构信息

Institute of Chinese Materia Medica, Academy of Chinese Medical Sciences, Beijing 100700, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2006 Dec;31(23):1940-3.

Abstract

OBJECTIVE

Searching and identifying SNP in Panax species and using multiplex allele-specific PCR (MAS-PCR) to authenticate P. ginseng and P. quenquefolium.

METHOD

Based on genbank database of Panax species, using DNAMAN to align the sequences, identify SNP of P. ginseng and P. quenquefolium. Design allele-specific primers for P. ginseng and P. quenquefolium, optimize the PCR reaction system including the usage amount of Taq, dNTP, primer, etc. Optimized system was performed with the total DNA of 20 different sources of P. ginseng and P. quenquefolium.

RESULT

When the annealing temperature was 66 'C, the template DNA of P. ginseng could be amplified 249 bp band whereas P. quenquefolium amplified 1 049 bp band.

CONCLUSION

The MAS-PCR have the advantages of highly specific, good reproducibility and could be identify P. ginseng and P. quenquefolium in the same PCR tube. It was a potential method to use in the molecular identification of other meteria medica.

摘要

目的

在人参属植物中搜索并鉴定单核苷酸多态性(SNP),并使用多重等位基因特异性PCR(MAS-PCR)对人参和西洋参进行鉴定。

方法

基于人参属植物的基因库数据库,使用DNAMAN软件对序列进行比对,鉴定人参和西洋参的SNP。设计人参和西洋参的等位基因特异性引物,优化PCR反应体系,包括Taq酶、dNTP、引物等的用量。使用20种不同来源的人参和西洋参的总DNA对优化后的体系进行检测。

结果

当退火温度为66℃时,人参的模板DNA可扩增出249 bp的条带,而西洋参可扩增出1049 bp的条带。

结论

MAS-PCR具有特异性高、重复性好的优点,可在同一PCR管中鉴定人参和西洋参。它是一种可用于其他中药材分子鉴定的潜在方法。

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