一种改进的Chelex-100法提取单头螨DNA

Extraction of DNA from single mite by promoted Chelex-100 method

提供了一种改进的Chelex-100法从单头螨中提取DNA,通过充分研磨以及反复冻融从单头螨得到的DNA可直接用于PCR。实验结果表明:与酚-氯仿法相比,此方法提取单头螨的DNA效果理想,操作简单,稳定性好;以3种不同类群螨为验证样,表明Chelex-100法可广泛用于各种螨的单头螨DNA提取;对于70%的酒精和奥氏螨保存液分别保存的个体,以及永久玻片标本,运用此方法均可提取出DNA,其中酒精保存标本提取效果最好;所提取的DNA在-20℃条件下可做长期保存。实验证明,改进的Chelex-100法是一种获取螨类等微小动物基因的较好方法。

Chelex-100 resin method was promoted to extract DNA from a single mite in this paper.The template obtained by Chelex-100 resin was tested by polymerase chain reaction followed with directly sequencing.The COI gene in mitochondrial DNA and the ITS gene from nuclear DNA were tested in five different mites.Compared with traditional method,the method was better and more stable.It might be suitable for all kinds of mites.It can be used to extract DNA from the single mite stored in different methods,such as 70% alcohol preservation,Oudmans＇ fluid preservation and berlese sgumchloral fluid preservation,which the first one was detected as the best one.And the template extracted by the method could be stored for a long time in the-20 ℃.To conclude,it showed that Chelex-100 method was suitable to extract DNA from tiny animals.