title: Illumina MiSeq sequencing; RNAseq profiling of a lpp1663 mutant compared to its isogenic parent Legionella pneumophila strain Paris
design: RNAseq profiling of a lpp1663 mutant compared to its isogenic parent Legionella pneumophila strain Paris
library_construction_protocol: Bacterial cultures were centrifuged at 5000g for 5 minutes. Pellets were stored at -80C Legionella pneumophila strains in this analysis were grown in AYE medium until they reach an optical density of 0.8 to 0.9 RNA samples were treated with DNAse I (Zymo Research) and further purified on silica-based column (Zymo Research) Depletion of ribosomal RNA with Ribo-Zero rRNA Removal Kit (Epicentre) RNA were extracted from frozen pellet by dissolving in 50l of RNAsnap buffer (18 mM EDTA, 0.025% SDS, 1% 2-mercaptoethanol, 95% formamide) and heated 7 minutes @ 95C. 1 mL of Trizol (Invitrogen) waas then added. RNA extraction was performed as recommended by the Trizol manufacturer. Library construction using TruSeq Stranded mRNA Library Prep Kit (Illumina)