RNA isolated from 3-day P. aeruginosa biofilms using QIAGEN RNeasy Mini Kit was subjected to Ambion TURBO DNA-free DNase Treatment. 2 ug of RNA was subjected to rRNA depletion using the Ambion MICROBExpress Bacterial mRNA Enrichment Kit. The  Ion Total RNA-Seq kit v2 was then used to  construct the cDNA library from the enriched mRNA.  The cDNA library was used to prepare  sequencing templates using the Ion PGM Template OT2 200 kit on the Ion OneTouch2 instrument.  Sequencing was carried out using Ion Sequencing 200 Kit v2 and the Ion 316v2 Chip on the Ion Torrent PGM insturment.