RNA-Seq of Neocallimastix frontalis 27 : glucose
All cultures were started from stock cultures stored in liquid nitrogen at the Agriculture and Agri-Food Canada Research Centre in Lethbridge Alberta Canada. Fungal biomass was produced under strict anaerobic conditions at 39 C in Lowe s semi-defined medium (Lowe et al., 1985) modified with 0.67% (wt/vol) of glucose. After 72 h of growth, fungal mycelia were harvested and immediately frozen in liquid nitrogen. Frozen mycelia were ground to a fine powder using a mortar and pestle and re-suspended in Trizol reagent according to the specifications of the manufacturer (Life Technologies Inc. Burlington, ON). Total RNA was isolated according to the manufacturer s instructions and further purified using the MEGAclear kit from Ambion (Life Technologies Inc. Burlington, ON). RNA quality was monitored using an Agilent Bioanalyzer 2100 by running samples on a RNA 6000 nano chip. After poly-A enrichment of mRNA, strand-specific RNA libraries were prepared and sequenced on an Illumina HiSeq 2000 instrument at the McGill University / Genome Quebec Innovation Centre.