Jiang Wenjun, Liang Peng, Wang Buying, Fang Jianhuo, Lang Jidong, Tian Geng, Jiang Jingkun, Zhu Ting F
School of Life Sciences, Center for Synthetic and Systems Biology, Ministry of Education Key Laboratory of Bioinformatics, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, China.
State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, State Environmental Protection Key Laboratory of Sources and Control of Air Pollution Complex, Tsinghua University, Beijing, China.
Nat Protoc. 2015 May;10(5):768-79. doi: 10.1038/nprot.2015.046. Epub 2015 Apr 23.
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.
宏基因组测序已广泛应用于研究来自土壤、海洋、沉积物和淡水等各种环境中的微生物群落。尽管如此，空气中微生物群落的宏基因组测序在技术上仍然具有挑战性，部分原因是可收集的大气颗粒物质量有限且所含生物成分较低。在此，我们提出了一种优化方案，可从收集的颗粒物中提取多达数十纳克的空气传播微生物基因组DNA。通过改进的测序文库制备方案，这一数量足以用于下游应用，例如对空气传播微生物群落中的各种基因进行宏基因组测序取样。所描述的方案在2至3天内需要约12小时的实验台操作时间，并且可以在实验室中使用标准分子生物学设备进行。该方案的修改版本也可用于从其他质量有限或生物成分较低的环境样品中提取基因组DNA。
