Optimized DNA extraction and metagenomic sequencing of airborne microbial communities - Nature Protocols, 10(5), p.768-779, 2015 .

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.


AIR POLLUTION
AIRBORNE MICROORGANISM
ARTICLE
CONTROLLED STUDY
DNA BINDING
DNA DETERMINATION
DNA EXTRACTION
DNA LIBRARY
DNA SEQUENCE
GENOME ANALYSIS
METAGENOMICS
MICROBIAL COMMUNITY
NONHUMAN
PARTICULATE MATTER
PRIORITY JOURNAL
PRIORITY JOURNAL
SEQUENCE ANALYSIS
BIOCHEMISTRY
CHINA
DEVICES
DNA SEQUENCE
ENVIRONMENTAL MONITORING
EQUIPMENT DESIGN
GENETICS
ISOLATION AND PURIFICATION
METAGENOME
MICROBIAL CONSORTIUM
MICROBIOLOGY
PROCEDURES
QUALITY CONTROL
WORKFLOW