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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 compare the method of preparation of total oral DNA from human oral saliva in different time periods. Methods: Phenol-chloroform extraction method was used to prepare oral salivary microbial total DNA at different time points in healthy population by QIAamp DNA Micro Kit. The total DNA extracted is identified using a UV spectrophotometer, PCR or the like. Results: The total DNA of oral microorganisms was successfully prepared by both methods. The total DNA extracted by different methods is different in fragment size, purity, yield and so on. There was no significant difference in oral salivary microbial total DNA in different time periods. CONCLUSION: Both methods of total DNA extraction can be used for metagenomics research. According to different experimental purposes, more suitable methods can be selected. Saliva from different donors of the same donor can be mixed for metagenomic studies in the short term.