title: RNAseq of Staphylococcus aureus MRSA N315
design: For RNA preparation, the strain was grown over night on solid LB media. A few single colonies were resuspended in 100 l RNAse free water and centrifuged for 3 min at 8,000  g. The pellet was resuspended in 100 l RNAse free water and vortexed for 3 min. A volume of 100 l of an 1:1 chloroform / phenol solution was added to the suspension and incubated for 30 min at 70C with shaking. The phases were separated at 10,000  g for 10 min and 100 l of the supernatant was removed and transferred to a fresh tube. The RNA extraction was prepared by Qiagen RNeasy Kit as described in the manufacturers instructions. 10 g total RNA per sample were used. Ribosomal RNA was depleted using the Ribo-Zero rRNA removal kit for Bacteria with subsequent purification of the rRNA-depleted sample by ethanol precipitation. Subsequently, a Illumina-compatible cDNA library was created using the TruSeq Stranded Total RNA Sample Prep Kit. The library was sequenced on an Illumina MiSeq sequencer in a 2  50 bp run.