GSM1666966: 10870ΔrocA; Streptococcus pyogenes; RNA-Seq
GAS cultures were incubated with RNAprotect™ (Qiagen) for >5 min at room temperature, pelleted and frozen at -80 °C overnight.  Cells were resuspended in RLT-BME and lysed using FastPrep Lysing Matrix B tubes (MP Biomedicals) in FastPrep-24 machine.  Samples were processed twice at speed 4 for 15 sec.  Total RNA was then isolated using miRNEasy Mini Kit (Qiagen) and eluted in nuclease-free water.  Off-column DNase treatments, using DNA-free DNase (Ambion), was performed at 37 °C.  The mRNA was then enriched using Ribo-Zero rRNA Removal Kit (Bacteria) to deplete rRNA contamination. All cDNA libraries were constructed using ScriptSeq™ RNA-Seq Library Preparation Kit (Epicentre) and processed according to manufacturer's protocol.
GEO Accession
GSM1666966