Analysis of the collective RNA pool from a microbial community - the metatranscriptome - yields valuable information on microbial gene expression patterns and biogeochemical processes in natural environments. Molecular and analytical tools for analyzing metatranscriptomes using high-throughput sequencing have advanced rapidly in recent years and continue to evolve and expand. The technique is increasingly available to individual research projects, even those with a modest budget or lacking an extensive bioinformatics toolkit. A core set of metatranscriptomic practices can now be identified, with key steps including RNA extraction, messenger RNA (mRNA) enrichment, synthesis of complementary DNA (cDNA), shotgun sequencing of cDNA, and bioinformatic analysis of sequence data. This chapter explores key questions that researchers should consider before beginning a metatranscriptomic study and then describes in detail the major steps of a sequencing-based metatranscriptomic analysis, from RNA isolation to functional and taxonomic analysis of sequence data. The questions and methods described here provide an introductory framework for environmental microbiologists interested in using metatranscriptome sequencing to explore microbial community gene expression.