真菌DNA提取方法的建立和比较  

Comparison of three methods of genomic DNA extraction from fungus

作　　者：杨艳秋[1] 王丽[2] 贺丹[2] 孙文通[2] 张宇[2] 横山耕治[3] 

机构地区：[1]吉林大学第一医院,吉林省长春市130021 [2]吉林大学基础医学院病原生物学教研室,吉林省长春市130021 [3]日本国立千叶大学真菌医学研究中心

出　　处：《中国组织工程研究与临床康复》2007年第50期10093-10096,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30571773);吉林省科技发展计划重点资助项目(20050205-3)~~

摘　　要：目的:DNA的提取效率,尤其是DNA的纯度严重影响实验结果,同一标本采用不同方法或不同标本采用相同方法提取DNA,其纯度和提取率有很大差异。以丝状真菌为材料.进行DNA提取.对真菌DNA的提取方法进行优化,探讨其最佳DNA提取方法方法:实验于2004-06/2006-12在吉林大学基础医学院真菌实验室进行以临床常见致病真菌标本为材料,采用玻璃珠-盐析法、CTAB法、Gene TLE^(TM)法3种不同方法提取DNA,通过琼脂糖凝胶电泳比较不同方法提取DNA的效率。结果:同等菌体量下,A_(260)和A_(280)的比值采用Gene TLE^(TM)法获得的样品最高,表明其蛋白质含量相对最少.3种提取的基因组DNA琼脂糖凝胶电泳图都在相应的多重PCR体系中扩增出目的片段,Gene TLE^(TM)DNA抽提法带型最清楚、明亮。结论:Gene TLE^(TM)法提取DNA效率高、质量好,操作简单.适于DNA的提取。AIM： The DNA extraction rate, especially the DNA purity, has a strong influence on the experiment results. There is a big difference of purity and extraction rate in extracting DNA either towards the same sample with the different method or towards the different samples with the same method. In this study, in order to find the best DNA extraction method, we used filamentous fungi as the sample and optimized the method. METHODS： This study was carried out at Mycology Lab of Basic Medical College of Jilin University from June 2004 to December 2006. Using clinical common pathomycete as the samples, we took three methods, including glass-bead salt fractionation, CTAB and Gene TLETM, to extract DNA and compared the extraction rate via agarose gel electrophoresis method. RESULTS： Under the same condition, the ratio between A260 and A280 of Gene TLE^TM was the highest, which presented the protein content was relatively low. Comparing the DNA agarose gel electrophoretogram made by the three methods, all the DNA can amplify the aim fragment in multi-PCR and the DNA extracted by Gene TLETM showed the best result. CONCLUSION： Gene TLE^TM is suit to extract DNA and has the advantages of high extraction rate, good quality and simple operation.

关 键 词：丝状真菌 基因组DNA提取 GENE TLE^TM法 

分 类 号：R519[医药卫生—内科学]