摘要
目的建立一种无设备快速30 s提取中药材DNA的方法。方法使用常规定性滤纸作为核酸吸附的载体,通过裂解液的裂解,释放核酸并吸附在滤纸上,利用洗脱液进行核酸纯化,即可完成不到30s的整个核酸提取过程,吸附核酸的滤纸可直接放入反应体系进行扩增。结果此方法可以成功提取不同药用部位的中药材核酸,提取得到的核酸经扩增验证,可获得与传统提取方法相一致的检测结果。结论此新型快速中药材DNA提取方法简单易行、速度快、成本低,可使核酸提取变得越来越大众化,不再局限于专业人员和实验室环境,为分子生药学的应用和发展提供了新思路。
Objective To build an equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA from Chinese medicinal materials in 30 s.Methods A new rapid DNA extraction method which was suitable for traditional Chinese medicine,with equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA from Chinese medicinal materials in 30 s was verified.The filter paper strip or disk for the adsorbed nucleic acid can be directly amplified by the reaction system.Results This method can extract the nucleic acid successfully.The extracted DNA was amplified,and the results were consistent with the traditional extraction methods.Conclusion The new rapid DNA extraction method is simple,fast,and low cost,which will make nucleic acid extraction become more and more popular,and no longer confined to professional and laboratory environment.This study provides a new idea for the application and development of molecular pharmacology.
作者
杨璐
吴文如
付菲
周华
吴小燕
黄祖波
安鑫
YANG Lu;WU Wen-ru;FU Fei;ZHOU Hua;WU Xiao-yan;HUANG Zu-bo;AN Xin(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;State Key Laboratory of Quality Research in Chinese Medicine,Macao University of Science and Technology,Macao 000853, China)
出处
《中草药》
CAS
CSCD
北大核心
2019年第2期502-509,共8页
Chinese Traditional and Herbal Drugs
基金
澳门科技大学中药质量研究国家重点实验室开放课题(MUST-SKL-2016-07)
国家中医药管理局全国中药特色技术传承人才培训项目(2015481601003)
广州中医药大学2018年校级大学生创新创业训练计划项目(201810572172)
关键词
快速提取
DNA提取
中药材
核酸
滤纸
rapid extraction
DNA extraction
Chinese medicinal materials
nucleic acid
filter paper
作者简介
杨璐(1993-),女,硕士研究生,从事中药分子鉴定研究。Tel:(020)39358296,E—mail:1084730732@qq.com;通信作者:吴文如(1976-),男,博士,副教授,硕士生导师,从事中药分子鉴定研究。Tel:(020)39358296,E-mail:wuwenm@gzucm.edu.cn.
