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Evaluation of Methods for the Extraction of DNA from Drinking Water Distribution System Biofilms

机译:饮用水分配系统生物膜中DNA提取方法的评价

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摘要

While drinking water biofilms have been characterized in various drinking water distribution systems (DWDS), little is known about the impact of different DNA extraction methods on the subsequent analysis of microbial communities in drinking water biofilms. Since different DNA extraction methods have been shown to affect the outcome of microbial community analysis in other environments, it is necessary to select a DNA extraction method prior to the application of molecular tools to characterize the complex microbial ecology of the DWDS. This study compared the quantity and quality of DNA yields from selected DWDS bacteria with different cell wall properties using five widely used DNA extraction methods. These were further selected and evaluated for their efficiency and reproducibility of DNA extraction from DWDS samples. Terminal restriction fragment length analysis and the 454 pyrosequencing technique were used to interpret the differences in microbial community structure and composition, respectively, from extracted DNA. Such assessments serve as a concrete step towards the determination of an optimal DNA extraction method for drinking water biofilms, which can then provide a reliable comparison of the meta-analysis results obtained in different laboratories.
机译:尽管在各种饮用水分配系统(DWDS)中已对饮用水生物膜进行了表征,但对不同DNA提取方法对饮用水生物膜中微生物群落后续分析的影响知之甚少。由于已显示出不同的DNA提取方法会影响其他环境中微生物群落分析的结果,因此有必要在应用分子工具表征DWDS的复杂微生物生态之前选择一种DNA提取方法。这项研究使用五种广泛使用的DNA提取方法比较了具有不同细胞壁特性的选定DWDS细菌的DNA产量和质量。从DWDS样品中进一步选择并评估其提取DNA的效率和可重复性。使用末端限制性片段长度分析和454焦磷酸测序技术分别解释了提取DNA中微生物群落结构和组成的差异。此类评估是确定饮用水生物膜的最佳DNA提取方法的具体步骤,然后可以对在不同实验室获得的荟萃分析结果进行可靠的比较。

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