摘要
建立了一个适合中药材川贝母的聚合酶链式反应-限制性内切酶(PCR-RFLP)的分子鉴定方法。以提取的DNA为模板进行PCR扩增和酶切反应,真品川贝母扩增产物含有Sam I酶切位点,在100~250 bp之间存在明确的2条酶切条带,而伪品贝母扩增序列中没有Sam I酶切位点不能被切出。此外,还基于PCR-RFLP技术针对川贝母真伪性进行了相对定量分析研究。结果表明,含量10%及以上的真品川贝母能被稳定检测出。
In this study,a molecular identification method of polymerase chain reaction restriction endonuclease(PCR-RFLP)was developed for identification of Fritillariae cirrhosae bulbus.The extracted DNA was used as the template for PCR amplification,and the amplification products were digested with endonuclease.Because Fritillariae cirrhosae bulbus has Sam I cut site,2 distinct bands between 100 bp to 250 bp could be observed.However,the pseudo Fritillaria could not be cut.Based on the PCR-RFLP technique,the relative quantitative analysis standardization of Fritillariae cirrhosae bulbus was also studied.The results showed that products containing 10%or more authentic Fritillariae cirrhosae bulbus could be detected.
作者
赵仲麟
常志远
袁超
李瑞歌
王成
王永
兰青阔
ZHAO Zhonglin;CHANG Zhiyuan;YUAN Chao;LI Ruige;WANG Cheng;WANG Yong;LAN Qingkuo(College of Sciences,Henan Agricultural University,Zhengzhou 450002,China;School of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua,321004,China;Institute of Tianjin Agriculture Quality Standard and Testing Technology,Tianjin 300381,China)
出处
《河南农业大学学报》
CAS
CSCD
北大核心
2018年第2期249-253,共5页
Journal of Henan Agricultural University
基金
国家自然科学基金项目(31100067)
关键词
川贝母
分子鉴定
PCR-RFLP
Fritillariae cirrhosae bulbus
molecular identification
PCR-RFLP
