GSM1519044: BL3; Staphylococcus aureus; miRNA-Seq
Total RNA was extracted from the NL and BL samples using a modified phenol chloroform extraction method as follows.  The pelleted cells were resuspended in RNAse free water and incubated with an equal volume of 1:1 phenol/chloroform.  Phases were separated after centrifugation, and RNA solution was centrifuged at 12,000 x g for 10 minutes. Two times the volume of isopropanol was added to the aqueous layer. The RNA was pelleted, and the pellet was washed with ethanol on ice and resuspended in Elution Solution (Ambion).  Both samples were treated with DNase Inactivation Reagent (Ambion).  Enrichment of the RNA was performed using the MICROBExpress Bacterial mRNA Enrichment Kit (Ambion) which involves using a modified capture hybridization involving magnetic beads to remove 16S and 23S ribosomal RNAs.  RNA was analyzed at the University of Oklahoma Science Center’s lab for Molecular Biology and Cytometry Research using Agilent Bioanalyzer 2100 to verify the concentration and purity of the mRNA. Illumina MiSeq RNA library Building (TruSeq)
GEO Accession
GSM1519044