title: RNA-Seq of L.fermentum U-21
design: Total RNA (500 ng) was used to prepare libraries. Ribosomal RNA was removed using the Ribo-Zero Plus rRNA Depletion Kit (Illumina). Libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (NEB). RNA purification was performed with RNA Clean XP magnetic particles and final library purification was performed by Agencourt AMPure XP (Beckman Coulter). The size and quality of the libraries were assessed using a highly sensitive DNA chip (Agilent Technologies). Library concentrations were measured with the Quant-iT DNA Assay Kit, High Sensitivity (Thermo Fisher Scientific). Libraries were mixed equimolarly and diluted to a final concentration of 12 pM. Ready libraries were sequenced on an Illumina HiSeq 2500 with 1% Phix (Illumina) added as a control.