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The choice of the DNA extraction method may influence the outcome of the soil microbial community structure analysis

机译:DNA提取方法的选择可能会影响土壤微生物群落结构分析的结果

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Abstract Metagenomics approaches and recent improvements in the next-generation sequencing methods, have become a method of choice in establishing a microbial population structure. Many commercial soil DNA extraction kits are available and due to their efficiency they are replacing traditional extraction protocols. However, differences in the physicochemical properties of soil samples require optimization of DNA extraction techniques for each sample separately. The aim of this study was to compare the efficiency, quality, and diversity of genetic material extracted with the use of commonly used kits. The comparative analysis of microbial community composition, displayed differences in microbial community structure depending on which kit was used. Statistical analysis indicated significant differences in recovery of the genetic material for 24 out of 32 analyzed phyla, and the most pronounced differences were seen for Actinobacteria . Also, diversity indexes and reproducibility of DNA extraction with the use of a given kit, varied among the tested methods. As the extraction protocol may influence the apparent structure of a microbial population, at the beginning of each project many extraction kits should be tested in order to choose one that would yield the most representative results and present the closest view to the actual structure of microbial population.
机译:摘要元基因组学方法和下一代测序方法的最新改进已成为建立微生物种群结构的一种选择方法。有许多商业的土壤DNA提取试剂盒,由于其高效性,它们正在取代传统的提取方案。但是,土壤样品理化特性的差异要求分别针对每个样品优化DNA提取技术。这项研究的目的是比较使用常用试剂盒提取的遗传物质的效率,质量和多样性。微生物群落组成的比较分析显示,微生物群落结构的差异取决于所使用的试剂盒。统计分析表明,在32种分析的门中,有24种的遗传物质回收率存在显着差异,其中放线菌的差异最为明显。同样,使用给定试剂盒进行DNA提取的多样性指标和可重复性在测试方法之间也有所不同。由于提取方案可能会影响微生物种群的表观结构,因此在每个项目开始时,应对许多提取试剂盒进行测试,以选择能够产生最具代表性的结果并提供最接近微生物种群实际结构的视图的试剂盒。

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