鸡肠道菌群总DNA三种提取方法的比较

目的比较不同方法提取鸡肠道菌群总DNA的差异,为分子方法分析肠道菌群组成提供质量较高的DNA模板。方法采用反复冻融法、酶裂解法和试剂盒法（E.N.Z.A Stool DNA Kit）来提取鸡肠道菌群的总DNA,并根据DNA浓度及纯度、16S DNA扩增产物和ERIC-PCR产物所反映的片段多态性4个指标,对这3种方法提取的DNA质量进行比较。结果3种方法均能提取DNA,所得DNA都可以用于16S DNA的扩增,但后2种方法所得DNA的ERIC-PCR结果能反映出更高的菌群多样性。结论试剂盒法和酶裂解法所提取的DNA质量好,适合用于肠道菌群的分子生态研究。

Comparison of three methods for total DNA extraction from intestinal microflora of the fowl

Objective To screen a method that could provide a higher quality DNA template for molecular analysis of composition of intestinal microflora. Method Repeated freeze-thaw method, lysozyme method and E.N.Z.A Stool DNA Kit method were used to extract total DNA from intestinal microflora of the fowl. The quality of DNA extracted by the three methods was compared based on the concentration, purity, 16S DNA PCR products and length polymorphism of ERIC-PCR products. Result Total DNA extracted by any of the three methods could be used to 16S DNA amplification. But the result of ERIC-PCR of DNA extracted by the last two methods showed more microflora diversities. Conclusion The quality of total DNA extracted by E.N.Z.A Stool DNA Kit method and lysozyme method were better than that of repeated freeze-thaw method, so can both be used to study molecular ecology of intestinal microflora.