蚊虫线粒体基因组DNA提取方法  

The extraction method of mosquito mitochondrial genome DNA

作　　者：邹依霖[1] 丁奕然[1] 罗钱春 陈斌[1] 

机构地区：[1]重庆师范大学昆虫与分子生物学研究所重庆市动物生物学重点实验室,重庆401331

出　　处：《中国媒介生物学及控制杂志》2015年第4期333-336,共4页Chinese Journal of Vector Biology and Control

基　　金："两江学者"计划专项经费;美国国立卫生研究院NIH项目(1R01AI095184);联合国IAEA项目(18268/R0);国家自然科学基金(31372265;31071968);重庆市科技攻关重点项目(CSTC2012GG-YYJSB80002);大学生创新创业训练计划国家级创新训练项目(201310637006)~~

摘　　要：目的优化和改进一套适用于蚊虫线粒体基因组DNA的提取方法。方法从动物线粒体基因组DNA提取的一般方法、步骤和原理出发,主要采用改进的高盐沉淀法,并注意去除核DNA的污染。用紫外分光光度计、琼脂糖凝胶电泳和线粒体COⅠ基因PCR扩增产物鉴定所提的线粒体基因组DNA。结果改进后的方法适用于单只蚊虫,用凝胶电泳检测后发现无核DNA与蛋白质污染。改进后的方法简便易行,经过多种蚊虫线粒体基因组提取的实验表明,提取的蚊虫线粒体基因组DNA质量高,能满足PCR、克隆、测序分析等要求。结论优化和改进的提取方法简便易行,得到的蚊虫线粒体基因组DNA质量高,能满足相关分析要求。Objective To optimize the method for the extraction of mosquito mitochondrial genome DNA. Methods The optimized method was improved from the protocol for the extraction of animal mitochondrial genomes, and was based on the modification for high-concentration-salt precipitation and nuclear DNA removing. The mtDNA samples were tested using spectrophotometry, agarose gel electrophoresis and amplification products of CO I. Results The method was showed to be rapid and efficient for single mosquito in our studies. The gel electrophoresis demonstrated that the mtDNA samples extracted were essentially free of nuclear DNA and protein. A number of applications with different mosquito species indicated that the method was highly suitable to extract the mitochondrial genome DNA of individual mosquitoes. Conclusion The optimized method is easy and efficient for mosquito samples, and the extracted mtDNA is high quality and suitable to studies on mitochondrial DNA.

关 键 词：蚊科 线粒体基因组 DNA提取方法 差速离心 

分 类 号：Q966[生物学—昆虫学]