GSM4859681: Untreated_rep1; Escherichia coli; RNA-Seq
Cultures were treated with RNAprotect. Bacterial cultured were lyzed and RNA was extracted using the PureLink RNA Mini Kit (Ambion life technologies, Invitrogen, Carlsbad, California, USA), treated with DNAse and cleaned using the RNA Clean & Concentrator kit (Zymo Research, Irvine, CA, USA). The quality and the quantity of each RNA samples was analyzed using a bioanalyzer and the Qubit technology. Dual-barcode indexed RNA-seq libraries were generated from 50 ng total RNA each, using the Kapa Hyper Stranded RNA-seq kit (KapaBiosystems/Roche, Basel, Switzerland) following the instructions of the manufacturer. Libraries were analyzed via micro-capillary gel electrophoresis on a LabChip GX system (PerkinElmer, Waltham, MA), quantified by fluorometry on a Qubit instrument (LifeTechnologies, Carlsbad, CA) and pooled in equimolar ratios. The pool was quantified by qPCR with a Kapa Library Quant kit (KapaBiosystems/Roche) and sequenced on one lane of an Illumina HiSeq 4000 (Illumina, San Diego, CA) with single-end 100 bp reads.