穿山甲药材DNA快速提取方法研究  

Study on Rapid Extraction of DNA from Manis squama Commodities

作　　者：杨志业 刘文斌[2] 谭颖仪 刘昂 方竑 林锦锋[1] Yang Zhiye;Liu Wenbin;Tan Yingyi;Liu Ang;Fang Hong;Lin Jinfeng(Guangdong Institute for Durg Control,Guangzhou 510663;No.1 Substation of Guangzhou Institute for Drug Control,Guangzhou 510130,China)

机构地区：[1]广东省药品检验所中药室,广东广州510663 [2]广州市药品检验所一分所,广东广州510130

出　　处：《广东化工》2020年第11期35-36,51,共3页Guangdong Chemical Industry

基　　金：广东省中医药局科研项目(项目编号:20171040);广东省食品药品监督管理局科技创新立项资助项目(项目编号:2018TDZ02);国家药品监督管理局药品快速检验技术重点实验室资助(国药监科外函[2019]82号-24)。

摘　　要：目的建立穿山甲药材DNA快速提取方法。方法采用二硫苏糖醇(DTT)结合商用动物组织DNA提取试剂盒提取穿山甲药材中的基因组。考察取样量、DTT用量、裂解时间等对穿山甲药材中DNA提取质量的影响。结果最低取样量为5mg,DTT用量为100μL,裂解时间为30 min,细胞裂解完全,所有样品的DNA均可满足PCR要求。结论建立的DNA提取方法提取时间为40 min,快速便捷,取样量少,DNA提取完全,可为穿山甲药材DNA鉴别方法开发提供研究基础。Objective To establish the rapid method of DNA extraction for Manis squama.Methods Dithiothreitol(DTT)was applied with commercial animal tissue genomic DNA extraction kits to extract total DNA from Manis squama.The effects of sampling amount,DTT dosage,and lysis time on the quality of DNA were also investigated.Results The lowest sampling amount was 5 mg,DTT dosage was 100μL,the least lysis time was 30 min,scale cells can be lysised completely,and the quality of DNA of all samples can meet the requirements of PCR.Conclusion The extraction method could be finished in 40 min,and could support study basis for DNA identification of Manis squama because of fast,less sample requiring and complete DNA extraction.

关 键 词：穿山甲 DNA快速提取 二硫苏糖醇(DTT) 

分 类 号：TQ[化学工程]