Hu Ang, Lu Yahai, Hernández García Marcela, Dumont Marc G
College of Resources and Environment, Hunan Agricultural University, Changsha, China.
College of Urban and Environmental Sciences, Peking University, Beijing, China.
Methods Mol Biol. 2019;2046:163-174. doi: 10.1007/978-1-4939-9721-3_13.
Metatranscriptomics is a powerful tool for capturing gene expression patterns in microbial communities and investigating their responses to environmental conditions. Stable isotope probing (SIP) is a method to target specific functional groups of microorganisms in environmental samples. The combination of RNA-SIP with metatranscriptomic analysis enhances the detection and identification of mRNA from target microorganisms. In this chapter we provide a protocol for RNA-SIP, mRNA enrichment, and mRNA preparation for high-throughput sequencing using an example of targeting methanotrophs in rice field soil.
宏转录组学是一种用于捕捉微生物群落中基因表达模式并研究其对环境条件响应的强大工具。稳定同位素探测（SIP）是一种针对环境样品中特定功能微生物群的方法。RNA-SIP与宏转录组分析相结合可增强对目标微生物mRNA的检测和鉴定。在本章中，我们以稻田土壤中甲烷氧化菌为目标为例，提供了一种用于RNA-SIP、mRNA富集和mRNA制备以进行高通量测序的方案。
