一种水环境eDNA提取方法的建立  

Establishment of a Method for Environmental DNA Extraction from Water Samples

作　　者：黎慧 阚霞 魏宁 邵晨[1,2] 王宇[1,3] LI Hui;KAN Xia;WEI Ning(College of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua,Zhejiang 321004)

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江师范大学初阳学院,浙江金华321004 [3]浙江师范大学野生动物生物技术与保护利用浙江省重点实验室,浙江金华321004

出　　处：《安徽农业科学》2019年第9期108-110,115,共4页Journal of Anhui Agricultural Sciences

基　　金：浙江省自然科学基金项目(LY19C030005);浙江师范大学实验技术开发项目(SJ201815);2017年浙江省大学生科技创新活动计划暨新苗人才计划项目(2017R404060);浙江省野生动物生物技术与保护利用重点实验室2018年开放项目

摘　　要：[目的]建立高效实用的水样环境DNA提取方案。[方法]采用滤膜法和沉淀法对2个不同大小的水域进行eDNA提取并对所获得的eDNA进行PCR扩增。其中,滤膜法设置50、100、200 mL 3个水样体积试验组,每组3个平行;沉淀法设置10、20、40 mL 3个水样体积试验组,每组3个平行。[结果]大型水塘水样经滤膜法处理后获得的eDNA浓度更高;小型水池采用滤膜法和大型水塘采用沉淀法处理水样后经PCR检测可获得更明显的扩增结果。[结论]该研究建立的滤膜法和沉淀法可用于不同水环境eDNA的提取。[Objective] To establish an efficient and practical method for extracting environmental DNA (eDNA) from water sample.[Method]Using the filter membrane or precipitation method to extract eDNA from two different sizes of water body and conducting PCR amplification to detect the quality and quantity of eDNA.Four groups were set with different water volume 50,100,200 mL and negative control in filter membrane method.And the other four groups with water samples 10,20,40 mL and negative control were set in precipitation method.All groups have three parallel samples.[Result]The concentration of eDNA extracted by the filter membrane method from the large water body was higher than others.The eDNA from small and large water body handled by the filter membrane and precipitation methods respectively can be detected significantly by PCR amplification.[Conclusion]The methods used in this study can be applied to eDNA extraction in different water bodies.

关 键 词：环境DNA提取 水环境 滤膜法 沉淀法 

分 类 号：S181[农业科学—农业基础科学]