碱裂解法快速提取虫草属真菌DNA研究  

Rapid Extraction of DNA from Cordyceps by Alkaline lysis

作　　者：王茜[1] 侯飞侠[1] 王艺璇[1] 叶茂[1] 彭成[1] 沈才洪 国锦琳[1,2] 

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省部共建国家重点实验室培育基地,四川成都611137 [2]泸州老窖博士后科研工作站,四川泸州646606

出　　处：《时珍国医国药》2016年第4期995-997,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30801522;No.81373920;No.81541167);四川省中医药管理局课题(No.2012-F-019);四川省成都市科技计划(No.12DXYB185JH-002);国家固体酿造工程技术中心资助

摘　　要：目的探索一种适用于虫草属基因组DNA快速提取的方法。方法在无液氮条件下使用碱裂解法提取虫草DNA,使用ITS通用引物进行PCR扩增。考察了提取液配方、煮沸时间对DNA提取的影响以及中和剂Tris-HCl用量对PCR扩增的影响。利用优化的提取方法提取了虫草属5种虫草并用ITS通用引物进行PCR扩增。结果 80μl 0.5 mol/L Na OH进行提取,加入160μl 0.1 mol/L Tris-HCl中和,离心即可在10min内提取出虫草属真菌的DNA,PCR扩增能获得清晰目的条带。结论优化的碱裂解法可用于虫草属DNA的快速提取。Objective To develop a rapid method to extract DNA from Cordyceps. Methods The alkaline lysis buffer and the neutralizing buffer were used for rapid extraction of DNA from Cordyceps without liquid nitrogen. ITS primers were used to amplify PCR products for Cordyceps. The formula composition of extracting solution,the volume of neutralizing buffer and the time of lysis were investigated. Results The results showed that 80μL the buffer of 0. 5M sodium hydroxide with 160μL the buffer of Tris-HCl can extract DNA from Cordyceps,the PCR products from Cordyceps materials DNAs were obtained. Conclusion The optimized alkaline lysis method is suitable to extract DNA from Cordyceps.

关 键 词：虫草属 DNA快速提取 碱裂解法 PCR 

分 类 号：R282.6[医药卫生—中药学]