滤纸法快速提取中药材DNA方法的研究  

Rapid Extraction of DNA from Chinese Herbal Medicines by Filter Paper Method

作　　者：王沙沙 王芳[1] 张涵 侯飞侠[1] 国锦琳[1] WANG Sha-sha;WANG Fang;ZHANG Han;HOU Fei-xia;GUO Jin-lin(The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine,Key Laboratory of Systematic Research,Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学,中药材标准化教育部重点实验室,中药资源系统研究与开发利用国家重点实验室培育基地,四川成都611137

出　　处：《中国现代中药》2020年第2期213-218,共6页Modern Chinese Medicine

基　　金：国家自然科学基金资助(81872959,81373920,30801522);四川省青年科技创新团队项目资助(19CXTD0055)。

摘　　要：目的:为了建立一种快速提取中药材DNA的新方法,本研究考察了滤纸法对中药材DNA提取的适用性。方法:提取了33个中药材DNA,动物药使用COI(除了海马)、植物药使用ITS2、真菌使用ITS通用引物进行聚合酶链式反应(PCR)扩增。结果:通过琼脂糖凝胶电泳检测发现,27个中药材PCR扩增成功,扩增成功率达到81.8%。结论:研究结果表明滤纸法对于动物类、叶类、花类、全草类、果实类及根茎类药材DNA提取有较好的效果,提取时间可控制在30 s之内。滤纸法缩短了DNA提取时间,减少了提取成本,将在中药材分子鉴别中发挥重要作用。Objective:In order to establish a new method for rapid extraction of DNA from Chinese medicinal materials,the applicability of filter paper method to the extraction of DNA from Chinese medicinal materials was researched in this study.Methods:Total DNA of 33 Chinese medicinal materials were extracted.COI primer(except for Hippocampus),ITS2 and ITS primers were used to amplify animal,botanical and fungi medicine,respectively.Results:Agarose gel electrophoresis detection indicated 27 Chinese herbal medicines were amplified successfully and the success rate of amplification reached 81.8%.Conclusion:The results showed that the filter paper method had a good effect on the DNA extraction from leaves,flowers,whole grass,fruit,rhizome and animal medicines,and the extraction time could be controlled in 30 s.The filter paper method could greatly shorten DNA extraction time and reduce cost.It will play an important role in the field application of molecular identification of Chinese medicinal materials.

关 键 词：滤纸法 中药材 DNA提取 分子鉴定 

分 类 号：R282[医药卫生—中药学] R931.5[医药卫生—中医学]