一种水环境eDNA提取方法的建立

Establishment of a Method for Environmental DNA Extraction from Water Samples

[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.