GSM6858283: 2_2_750pq__2; Escherichia coli str. K-12 substr. MG1655; RNA-Seq
3 mL of induced culture was added to 6 mL of RNAProtect Bacteria Reagent (Qiagen) and vortexed, then left at room temperature to incubate for 5 minutes. Cells were pelleted, resuspended in 400 μL elution buffer, and then split into two tubes with one kept as a spare. One pellet was then lysed enzymatically with addition of lysozyme, proteinase-K, and 20% SDS. SUPERase-In was added to maintain the integrity of the RNA. RNA isolation was then performed according to  the RNeasy Mini Kit (Qiagen) protocol.  rRNA was depleted using the Ribo-Zero rRNA Removal Kit for gram negative bacteria according to the protocol. Libraries were constructed for paired-end sequencing using a KAPA RNAseq Library Preparation kit.