口腔微生物宏基因组学总DNA制备的方法学初探

An attempt to prepare genomic DNA for human oral microbial metagenome

目的:比较不同时间段人类口腔唾液微生物总DNA制备方法。方法:采用酚氯仿抽提法、QIAamp DNA Micro Kit法制备健康人群不同时间段口腔唾液微生物总DNA。使用紫外分光光度计,PCR等鉴定提取的总DNA。结果:两种方法均成功制备了口腔微生物总DNA。不同方法提取的总DNA在片段大小,纯度,得率等方面存在差异。短期内不同时间段口腔唾液微生物总DNA无显著差异。结论:两种总DNA提取方法均可用于宏基因组学研究,可根据不同的实验目的,选择更适合的方法。同一供体短期内不同时间段的唾液可混合用于宏基因组学研究。

Objective：To analysis of two methods for the extraction of microbial genomic DNA from different-period human saliva.Method：The two DNA extraction methods,traditional extraction method and QIAamp DNA Micro kit method,isolate the total microbial DNA from different-period human saliva.The two methods were evaluated comprehensively for the quantity,purity and length of the DNA and the microbial diversity.Result：The total DNA,extracting by the two methods,can be successfully applied to the PCR amplification.The total DNA by traditional method is small fragments,high impurity,but high concentration.The total DNA by QIAamp DNA Micro Kit is low concentration,but high purity,large segment.It showed that DNA extracted from human saliva was no significant difference along the three different-period groups.Conclusion：The two methods can both use for metagenome study,but should choose for the different study objective.The different-period human saliva during a short time can mix for the study of human oral metagenome.