Sample processing and cDNA preparation for microbial metatranscriptomics in complex soil communities.
Soil presents one of the most complex environments for microbial communities as it provides many microhabitats that allow coexistence of thousands of species with important ecosystem functions. These include biomass and nutrient cycling, mineralization, and detoxification. Culture-independent DNA-based methods, such as metagenomics, have revealed operational taxonomic units that suggest a high diversity of microbial species and associated functions in soil. An emerging but technically challenging area to profile the functions of microorganisms and their activities is mRNA-based metatranscriptomics. Here, we describe issues and important considerations of soil sample processing and cDNA preparation for metatranscriptomics from bacteria and archaea and provide a set of methods that can be used in the required experimental steps.
School of Agriculture and Food Sciences, The University of Queensland, Brisbane, Queensland, Australia.
Lilia C
LC
Carvalhais
Peer M
PM
Schenk
D001105:Archaea / Q000235:genetics; D001419:Bacteria / Q000235:genetics; D018076:DNA, Complementary / Q000302:isolation & purification*; D017753:Ecosystem; D020869:Gene Expression Profiling; D054892:Metagenome; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
D018076:DNA, Complementary; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S
Metatranscriptomics; RNA-seq; Sample processing; Soil; cDNA synthesis