We propagated H. somni 2336 on three TSA-blood plates (with 5% sheep red blood cells) for 16 hr or until a fresh lawn of cells was visible. IBC approval was not required for acquiring the plates as they were purchased through a commercial vendor: Fisher Scientific (Pittsburgh, PA), and manufactured by Becton Dickinson Diagnostic Systems, (Franklin Lakes, NJ). We washed the plates with brain heart infusion (BHI) broth, adjusted the culture to an OD620 nm = 0.8, and supplemented with RNAprotect reagent. The cells were harvested by centrifugation and stored at 280uC. We extracted total RNA using the RNeasy mini kit (Qiagen, Valencia, CA) following the manufacturer's protocol. Total RNA was treated with RNase-free DNAse (Invitrogen, Carlsbad, CA). Using Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA), we determined the RNA integrity number (RIN) of total RNA to be greater than 8. MICROBExpress TM Kit (Ambion, TX, USA), which specifically removes rRNAs, was used for mRNA enrichment. Small RNAs (i.e., tRNA and 5S rRNA) are not removed with this enrichment step (confirmed by Bioanalyzer).