GSM2754493: LT2 ridA Rep 2; Salmonella enterica subsp. enterica serovar Typhimurium str. LT2; RNA-Seq
Each 5 mL sample was quickly  centrifuged at 16,000 x g in 1.5 mL Eppendorf tubes, supernatant removed and pellets flash-frozen in liquid nitrogen and kept on dry ice.  Total RNA was extracted using the RNAsnapTM method (Stead, et al., 2012).  RNA was subjected to an ethanol precipitation and DNA/RNA pellets were resuspended in 90 μl UltraPureTM water at 4°C overnight prior to treatment with RNase-free Turbo DNase (Ambion), and being precipitated once more by sodium acetate-ethanol treatment. Resulting RNA was resuspended at 4°C for 3 h and stored at -80°C until use. The total RNA was extracted from four independent replicates of S. enterica wild-type and ridA mutant strains. In each of the resulting eight samples, rRNA was removed from 5 μg total RNA using the Ribo-Zero rRNA removal kit (Bacteria; Illumina) following manufacturer’s recommendations. A cDNA library was created from samples containing <10% rRNA contamination, as measured by the RNA 6000 pico kit for the Agilent 2100 bioanalyzer, and sequenced by the University of Georgia Genomics Facility (GGF) from these RNA samples. Specifically, one hundred nanograms of the rRNA-free RNA was used to create each sequencing library using the KAPA stranded RNA-seq kit (KAPA Biosystems).
GEO Accession
GSM2754493