title: Quantitative microbial metatranscriptomics.
abstract: The direct retrieval and sequencing of environmental RNA is emerging as a powerful technique to elucidate the in situ activities of microbial communities. Here we provide a metatranscriptomic protocol describing environmental sample collection, rRNA depletion, mRNA amplification, cDNA synthesis, and bioinformatic analysis. In addition, the preparation of internal RNA standards and their addition to the sample are discussed, providing a method by which transcript numbers can be expressed as absolute abundances in the environment and more readily compared to other biogeochemical and ecological measurements.
authors: Department of Marine Sciences, University of Georgia, Athens, GA, USA.
authors: Scott
authors: S
authors: Gifford
authors: Brandon
authors: B
authors: Satinsky
authors: Mary Ann
authors: MA
authors: Moran
mesh_terms: D019295:Computational Biology / Q000379:methods; D004783:Environmental Microbiology; D020869:Gene Expression Profiling / Q000379:methods; D056186:Metagenomics / Q000379:methods; D059467:Transcriptome