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An Improved Method for Soil DNA Extraction to Study the Microbial Assortment within Rhizospheric Region

机译:一种改进的土壤DNA提取方法以研究根际区域内的微生物分类

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摘要

The need for identification of soil microbial community mainly depends on direct extraction of DNA from soil, a multifaceted environment that is a major pool for microbial genetic diversity. The soil DNA extraction procedures usually suffer from two major problems, namely, inappropriate rupturing of cells and contamination with humic substances. In the present study, five protocols for single type of rhizospheric soil were investigated and their comparison indicated that the inclusion of 120 mM phosphate buffered saline (PBS) for washing and mannitol in the lysis buffer allowed the processing of soil sample in minimal time with no specific equipment requirement. Furthermore, DNA purity and yield were also improved, which allowed the exploitation of genetic potential of soil microbes within soil sample thereby facilitating the amplification of metagenomic DNA. The effectiveness of methods was analyzed using random amplification of polymorphic DNA. The banding patterns revealed that both the abundance and the composition of indigenous microbial community depend on the DNA recovery method.
机译:鉴定土壤微生物群落的需要主要取决于从土壤中直接提取DNA,土壤是一个多方面的环境,是微生物遗传多样性的主要来源。土壤DNA提取程序通常会遇到两个主要问题,即不适当的细胞破裂和腐殖质的污染。在本研究中,研究了针对单一类型根际土壤的五种方案,它们的比较表明,在裂解缓冲液中加入用于洗涤和甘露醇的120 phosphatemM磷酸盐缓冲盐水(PBS)可以在最短的时间内处理土壤样品,而无需特定设备要求。此外,DNA的纯度和产量也得到了改善,从而可以利用土壤微生物在土壤样品中的遗传潜力,从而促进宏基因组DNA的扩增。使用多态性DNA的随机扩增分析了方法的有效性。条带化模式显示,土著微生物群落的丰度和组成均取决于DNA的回收方法。

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