GSM4156711: m-Tyr 60 min treatment Rep 1; Escherichia coli; RNA-Seq
RNA purification was performed using the Qiagen®  RNeasy® Mini Kit.  One-ml of culture was removed for each time point and mixed with 2 ml of RNAprotect® Bacterial Reagent.  RNA was then purified accound to the Qiagen® protocols for “Enzymatic Lysis of Bacteria” and “Purification of Total RNA from Bacterial Lysate Using the RNeasy® Mini Kit”. Ribosomal RNA was removed from the RNA samples using the Thermo Fisher Scientific RiboMinus™ Transcriptome Isolation Kit for bacteria.  The resulting rRNA-depleted samples were then concentrated using the Qiagen® RNeasy® columns using the “RNA Clean Up” protocol. cDNA libraries were then created using New England Biolabs' (Ipswich, MA, USA) NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® and the Primer Set 1 of NEBNext® Multiplex Oligos for Illumina (Ipswich, MA, USA). Sequencing of the 12 cDNA libraries was performed using the Illumina MiSeq Personal Sequencer System with paired end 150 bp reads.