title: RNA-seq of Caenibius tardaugens: Progesterone replicates 1 to 3
design: Total RNA of C. tardaugens cells was extracted from cultures grown in minimal medium with 20 mM CDX and progesterone as carbon source. Cells where harvested in mid exponential phase (OD600 0.6) and stored at 80 C. Pellets where thawed and cells were lysed in 400 L TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5) containing lysozyme (50 mg/mL) following three freezingthawing cycles. High Pure Isolation Kit (Roche), followed by DNA-free DNA Removal Kit (Invitrogen) treatment, was used to obtained pure RNA. Purity and concentration were measured in a ND1000 spectrophotometer (Nanodrop Technologies). Ribosomal RNA was removed from the total RNA with Ribo-Zero rRNA Removal Kit to later construct a 100 bp paired-end library using TruSeq RNA Sample Prep Kit v2 that was quality-checked in an Agilent Technologies 2100 Bioanalyzer using a DNA 1000 chip. Library sequencing was performed in a HiSeq 3000 4000 (Illumina) using TruSeq 3000 4000 SBS Kit v3 as reagent.