一种高效的哺乳动物粪便DNA提取通用方法  

An Effective Method with High Currency for DNA Extraction from Mammalian Faeces

作　　者：赵健元[1,2,3] 李进华[1,2] 

机构地区：[1]安徽大学生命科学学院,安徽合肥230039 [2]安徽大学安徽省生态工程与生物技术重点实验室,安徽合肥230039 [3]安徽省体育科学技术研究所,安徽合肥230001

出　　处：《激光生物学报》2008年第5期695-700,646,共7页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30570237;30770288);教育部骨干教师资助计划(18557);安徽省教育厅科技创新团队计划项目(TD200703);安徽省优秀青年基金项目(040443070);安徽省人才开发基金项目;安徽大学学术创新团队项目资助

摘　　要：以粪便为材料提取动物DNA进行动物保护遗传学和分子生态学研究的关键是能否提取到高质量的粪便DNA。然而提取方法通用性不好和产物质量不高等问题阻碍了粪便DNA分析技术的推广。本文介绍的改进型十六烷基三甲基溴化铵提取法可广泛适用于各食性哺乳动物粪便DNA提取,在11种不同食性动物的粪便DNA提取实验中验证了它的可靠性和通用性。本方法成本低廉(3元/样),用实验室常规试剂即可完成粪便DNA提取,其产物纯度高于专用试剂盒QIAamp DNA Stool Kit,在拥有超过专业试剂盒提取效果的同时尽可能的降低了实验成本,有利于粪便DNA技术的推广。As an ideal noninvasive sampling way, extracting DNA from faecal sample alleviates sampling problem which existed longtermly in conservative genetics and molecular ecology studies. The core of this technology is whether high quality faecal DNA can be extracted from faecal samples. Although several methods had been developed at home and abroad, two problems below have not been solved yet. For one thing, most of the current methods are not good in different experiments. The researchers always got different amplification success rate between different animal species and individuals. For another, the quality of the DNA is not good enough for PCR amplification. These questions hampered the development and spread of molecular scatology badly. The ＂Improved CTAB method＂ introduced in this paper is applicable to many kinds of mammals no matter they have the same diet habit or not. With this method, high amphcation success rate had been obtained. In the validation of the new method＇ s effectiveness, high quality and quantity DNA had been extracted from 11 different kinds of mammals. The results of sequencing confirmed the reliability and currency of the new method. Although the ＂QIAamp DNA stool kit＂ has good currency in faecal DNA extracting, but the high price restrict the far-ranging application of the kit. In contrast to that kit, the cost of ＂improved CTAB method＂ is very cheap.And the quality of DNA extracted by improved CTAB method is much better than the DNA obtained by that kit. Therefore, the new method actualizes the currency of extracting method. It will facilitate the spread of molecular scatology.

关 键 词：粪便 DNA提取 通用性 CTAB 十六烷基三甲基溴化铵 

分 类 号：Q78[生物学—分子生物学]