Bistable Expression of CsgD in Salmonella Connects Virulence to Persistence
RNA was treated with the MICROBExpress™ Bacterial mRNA Enrichment Kit (Life Technologies #AM1905) prior to preparing cDNA. First strand cDNA was synthesized from RNA using random hexamer priming and reverse transcriptase. Second strand cDNA synthesis was performed with DNA polymerase I in presence of dATP, dGTP, dCTP and dUTP. Size selection was 70-200 bp before ligation of Illumina adapters and final fragment sizes were approximately 196 to 326 bp. The second cDNA strand containing UTP was selectively removed by uridine digestion prior to library amplification and sequencing.