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Indirect DNA extraction method suitable for acidic soil with high clay content

机译：适用于粘土含量高的酸性土壤的间接DNA提取方法

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class="kwd-title">Method name: Indirect DNA extraction class="kwd-title">Keywords: Microbial community, Extracellular DNA, Acid sulfate soil, 16S rRNA gene class="head no_bottom_margin" id="abs0010title">AbstractDNA extraction is an essential procedure when investigating microbial communities in environmental samples by sequencing technologies. High clay soils can be problematic as DNA adsorbs to the clay particles and can thereby be preserved from lysed, non-viable cells for a substantial period of time. In order to accurately estimate the intact and living microbial community in the soil, extracellular DNA from dead, remnant bacterial cells needs to be removed prior to DNA extraction. One possibility is to use a sodium phosphate buffer to release both extracellular DNA and bacterial cells from the clay particles. After removing the extracellular DNA by centrifugation, the remaining viable cells can be harvested and DNA extracted. The described method is a modification of a procedure for separating extracellular DNA and bacterial cells from acidic clay soils. class="first-line-outdent" id="lis0005">
  • • The modified method increases bacterial cell yields from acidic clay soils, such as acid sulfate soil.
  • • The modified method eliminates some steps from the original method, as only DNA from intact bacterial cells is required.
  • • The indirect DNA extraction method increases the workload compared to standard direct extraction methods, but subsequent downstream analyses will give a more representative picture of the viable microbial community composition in the soil.
    • 机译：<！-fig ft0-> <！-fig @ position =“ anchor” mode =文章f4-> <！-fig mode =“ anchred” f5-> <！-fig / graphic | fig / alternatives / graphic mode =“ anchored” m1-> class =“ kwd-title”>方法名称：间接DNA提取 class =“ kwd-title”>关键字：微生物群落，细胞外DNA，酸性硫酸盐土壤，16S rRNA基因 class =“ head no_bottom_margin” id =“ abs0010title”>摘要 DNA提取是通过测序技术研究环境样品中微生物群落的重要步骤。高黏土可能会成为问题，因为DNA会吸附到黏土颗粒上，因此可以在相当长的一段时间内不溶解的，没有活力的细胞中保存下来。为了准确估计土壤中完整和活的微生物群落，需要在提取DNA之前从死的残留细菌细胞中去除细胞外DNA。一种可能性是使用磷酸钠缓冲液从粘土颗粒中释放出细胞外DNA和细菌细胞。通过离心除去细胞外DNA后，可以收集剩余的活细胞并提取DNA。所描述的方法是从酸性黏土中分离细胞外DNA和细菌细胞的方法的改进。 class =“ first-line-outdent” id =“ lis0005”> <！-list-behavior =简单的前缀-word = mark-type = none max-label-size = 9->
  • •修改后的方法可以提高酸性粘土土壤（如酸性硫酸盐土壤）的细菌细胞产量。
  • •修改后的方法从原始方法中省去了一些步骤，因为只需要完整细菌细胞中的DNA。
  • •间接DNA提取方法与标准的直接提取方法，但随后的下游分析将对土壤中可行的微生物群落组成更具代表性。

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