Vettraino Anna Maria, Luchi Nicola
Department for Innovation in Biological, Agro-food and Forest Systems (DIBAF), University of Tuscia, Viterbo, Italy.
Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR), Sesto Fiorentino, Florence, Italy.
Methods Mol Biol. 2022;2536:103-107. doi: 10.1007/978-1-0716-2517-0_6.
Metagenomics offers the possibility to study the microbial community of soils at large scale, allowing the characterization also of unculturable microorganisms. Availability of high-quality DNA is a prerequisite of this approach. However, since soils have highly heterogeneous physicochemical properties, several parameters need to be considered for the development of the best molecular practices. A method for isolation of high-molecular-weight and good-quality metagenomic DNA from different soil samples is described here. The protocol combines physical and chemical strategies to ensure efficient cell lysis and precipitation of humic impurities-free DNA suitable for downstream processing for metagenomics study.
宏基因组学为大规模研究土壤微生物群落提供了可能性，使得不可培养微生物也能够得到表征。高质量DNA的可得性是这种方法的一个先决条件。然而，由于土壤具有高度异质的物理化学性质，为了开发最佳的分子方法，需要考虑几个参数。本文描述了一种从不同土壤样品中分离高分子量和高质量宏基因组DNA的方法。该方案结合了物理和化学策略，以确保有效的细胞裂解和沉淀不含腐殖质杂质的DNA，适合用于宏基因组学研究的下游处理。
