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Comparison of DNA extraction and purification methods from different soils for metagenomic sequencing

机译:不同土壤中DNA提取和纯化方法对分析测序的比较

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There is an increased interest in the extraction of nucleic acids from various environmental samples, since only a minority of naturally occurring microbes can be cultured using standard techniques. Nucleic acids extraction and purification from soils are extremely challenging due to the low biomass, high organic contents and high variability of soil types. This has been regarded as one of the major difficulties that hamper the development of soil microbial ecology study. No commercial nucleic acids kits currently available are capable of preparing the DNAs without modifications. The cost can be very high for DNA extraction from extreme environmental soil samples, such as soils that have extreme high or low pHs. In this work, we developed and optimized soil DNA extraction and purification methods on different soils and compared the impact of three different DNA extraction protocols on DNA yield and purity. For the three different types of soil we used, direct extraction obtained the highest DNA recover rate, but required more cleanup steps. MoBio PowerSoil~R DNA Isolation Kit yields less but do not require as many downstream cleaning steps. Both of the two methods obtained a more abundant microbial community than Meta-G-Nome~(TM) DNA Isolation Kit.
机译:对来自各种环境样品的核酸提取的兴趣增加,因为只有使用标准技术可以培养少数天然存在的微生物。由于生物量低,有机含量高,土壤类型的高可变性,核酸从土壤中提取和纯化是极其挑战性。这被认为是妨碍了土壤微生物生态学研究发展的主要困难之一。目前可用的商业核酸试剂盒能够在没有修饰的情况下制备DNA。对于从极端的环境土壤样​​品中的DNA提取,例如具有极高或低于pHS的土壤,成本非常高。在这项工作中,我们在不同土壤中开发和优化了土壤DNA提取和纯化方法,并比较了三种不同DNA提取方案对DNA产量和纯度的影响。对于我们使用的三种不同类型的土壤,直接提取获得了最高的DNA恢复率,但需要更多的清理步骤。 Mobio PowerSoil〜R DNA隔离套件产生较少但不需要多个下游清洁步骤。这两种方法中的两种比Meta-G-Nome〜(TM)DNA分离试剂盒获得了更丰富的微生物群落。

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