中国中药杂志

2014, v.39(19) 3668-3672

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快速PCR方法在金银花真伪鉴别中的应用
Rapid PCR authentication Lonicera japanica

蒋超;侯静怡;黄璐琦;袁媛;陈敏;金艳;
JIANG Chao;HOU Jing-yi;HUANG Lu-qi;YUAN Yuan;CHEN Min;JIN Yan;Institute of Resource Ecology and Traditional Chinese Medicine Resources,Beijing Normal University;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

摘要(Abstract):

建立简单快速的金银花真伪分子鉴别方法。该研究依据金银花trn L-trn F 625位G/T SNP位点设计快速位点特异性PCR引物,优化位点特异性PCR条件,对金银花及其9种混淆品进行扩增及检测。当在87℃预变性1 min;87℃变性5s,68℃延伸5 s,30个循环时,通过加入SYBR Green I染料染色,金银花样品显绿色荧光,混淆品无荧光。整个PCR反应可在30 min内完成。该研究结果说明快速位点特异性PCR能简单快速鉴别金银花及其混淆品。
To simply and rapid authenticate Lonicera japanica. Rapid allele-specific PCR primer was designed base on trn Ltrn F 625 G / T Single nucleotide polymorphism and the PCR reaction systems including annealing temperature was optimized; optimized results were performed to authenticate L. japanica and its 9 adulterants. When 100 × SYBR Green I was added in the PCR product of87 ℃ initial denatured 1 min; 87 ℃ denatured 5 s,68 ℃ annealing 5 s,30 cycle; L. japanica visualize strong green fluorescence under365 nm UV lamp whereas adulterants without. The results indicate rapid allele-specific PCR could authenticate L. japanica and its adulterants rapidly and simply.

关键词(KeyWords): 快速PCR;金银花;分子鉴定;荧光检测
rapid PCR;Lonicera japonica;molecular authentication;fluorescence

Abstract:

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基金项目(Foundation): 国家自然科学基金项目(81373959);; 中医药行业科研专项(201407003)

作者(Author): 蒋超;侯静怡;黄璐琦;袁媛;陈敏;金艳;
JIANG Chao;HOU Jing-yi;HUANG Lu-qi;YUAN Yuan;CHEN Min;JIN Yan;Institute of Resource Ecology and Traditional Chinese Medicine Resources,Beijing Normal University;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

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