孔雀粪样宏基因组DNA提取方法的优化

Optimization of Extraction Method of Metagenomic DNA from Pavo Fecal Samples

摘要: 目的：本研究旨在优化孔雀粪样中菌群基因组DNA的提取方法，以便为后续的研究奠定基础。方法：从野生动物园和圆通山动物园采集孔雀粪样，基于粪样用量、杂质去除方法和消化酶的添加等角度优化了孔雀粪样宏基因组DNA的提取方法，并通过16S rDNA基因和ERIC的PCR扩增验证了纯化DNA的效果。结果：采用0.5 g孔雀粪样、利用丙酮和乙醇交替漂洗样品及消化液中添加CTAB是提取孔雀肠道宏基因组DNA的优化关键点，通过该方法提取的DNA可用于后续的PCR扩增。结论：根据优化的方法可有效地提取孔雀粪样中的基因组DNA，研究结果为孔雀肠道菌群的研究奠定了基础。

Abstract: Objective: The aim of this study is to optimize the extraction method of genomic DNA of microbiota from peacock feces, so as to lay a foundation for follow-up researches. Methods: The dung samples from Pavo were collected from the Wildlife Park and Yuantongshan Zoo in Yunnan province. The extraction method of metagenomic DNA from peacock dung samples was optimized based on the sampling amount of dung, the methods of impurity removal and the addition situation of digestive enzymes, et al. The effect of DNA purification was verified by PCR amplification of 16S rDNA gene and ERIC. Results: The key points for the optimization of extracting metagenomic DNA from peacock gut were using 0.5 g peacock fecal sample, rinsing the sample alternately with acetone and ethanol and adding CTAB to the digestive fluid. The DNA extracted by this method could be used for subsequent PCR amplification. Conclusion: The genomic DNA could be effectively extracted from peacock feces using the optimized method in this study, and the results would have laid a foundation for the research of peacock intestinal microbiota.