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Optimized DNA extraction and metagenomic sequencing of airborne microbial communities

2015; Nature Portfolio; Volume: 10; Issue: 5 Linguagem: Inglês

10.1038/nprot.2015.046

1754-2189

Wenjun Jiang, Peng Liang, Buying Wang, Jianhuo Fang, Jidong Lang, Geng Tian, Jingkun Jiang, Ting Zhu,

Air Quality Monitoring and Forecasting

This protocol enables collection of airborne particulate matter; and after sample pretreatment, it allows sufficient quantities of microbial DNA to be extracted and prepared for downstream applications such as metagenomic sequencing. Metagenomic sequencing has been widely used for the study of microbial communities from various environments such as soil, ocean, sediment and fresh water. Nonetheless, metagenomic sequencing of microbial communities in the air remains technically challenging, partly owing to the limited mass of collectable atmospheric particulate matter and the low biological content it contains. Here we present an optimized protocol for extracting up to tens of nanograms of airborne microbial genomic DNA from collected particulate matter. With an improved sequencing library preparation protocol, this quantity is sufficient for downstream applications, such as metagenomic sequencing for sampling various genes from the airborne microbial community. The described protocol takes ∼12 h of bench time over 2–3 d, and it can be performed with standard molecular biology equipment in the laboratory. A modified version of this protocol may also be used for genomic DNA extraction from other environmental samples of limited mass or low biological content.