RNA-seq of Acinetobacter baylyi ADP1 TadA_D54E + mRFP expressed from the pBTK402 plasmid colony 4
Samples were grown in LB medium, each started from a single and different colony. RNA was extracted in mid-log phase at OD of 0.5-0.8. RNA was extracted using GeneJET RNA Purification Kit (Thermo Scientific #K0731). The RNA sample was treated with 4 units of DNase I (NEB # M0303L) for 20 minutes at 37 C. Finally, following incubation of 15 minutes at 65 C. Ribosomal RNA was depleted using NEBNext rRNA Depletion Kit (Bacteria) (New England Biolabs, # E7850). Libraries were constructed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, #E7760). Finally, RNA-seq libraries were sequenced on the NovaSeq X platform (Illumina).