Isolation of High Quality RNA for Metatranscriptomic Analysis of Lignocellulose Digestion in the Rumen.

ABSTRACT

Metatranscriptomics can be used to examine both the composition of a microbial community as well as its metabolic activity under a particular set of conditions and complement metagenomic studies. The availability of low-cost, high-throughput next-generation sequencing has led to a rapid increase in the number of metatranscriptomic studies being undertaken. One of the primary difficulties when conducting transcriptomics is the ability to isolate high-quality RNA from samples of interest. The application of metatranscriptomics in rumen microbiology is still relatively novel but there is a significant push toward applying this technology in this field. In this protocol, we outline the method that is used routinely in our laboratory to purify high quality RNA from rumen contents that are suitable for metatranscriptomic sequencing using RNA-seq.