Name: GSM8295805
Instrument: Illumina HiSeq 3000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Bacterial pellets were incubated in a cell wall lysis mixture TE buffer pH 8 (30 mMTris, 1mMEDTA, 15 mg/mL and 200μg/mL proteinase K) containing mutanolysin, lysostaphin and lysozyme at 37 °C for 30 minutes. TRK lysis buffer and 70% ethanol were added to each sample, prior to transferring to E.Z.N.A RNA isolation columns. RNA was isolated following the manufacturer's instructions and DNA was selectively degraded using the DNA-free DNA removal kit.The RNA was precipitated with 0.1 volume 3 M sodium acetate (ThermoFisher #S209) and 3 volumes of 100% ethanol, recovered by centrifugation and washed with ice cold 70% ethanol. A ribosomal RNA-depleted cDNA library was prepared according to the manufacturer's instructions using the Universal Prokaryotic RNA-Seq Prokaryotic AnyDeplete kit (NuGEN #0363-32) and sequenced with Illumina HiSeq. Raw base calls were converted to fatsq files using Bcl2fastqs. Filtered reads were aligned to the Newman (Refseq NC_009641.1) reference genome using STAR-Aligner v2.7.3a. The mapped reads were annotated for read groups and marked for duplicates using the Picard tools v2.22.3.