title: GSM8838473: MexTKO_OD2_1; Pseudomonas aeruginosa; RNA-Seq
library_construction_protocol: Wild-type PAO1 and PAO1∆mexT cultures were started from single colonies in LB-MOPS in biological triplicate. Cultures were incubated for 18 hours at 37˚C with shaking. Cultures were back diluted to an OD600 of 0.1 until cultures doubled, and then cultures are diluted back to an OD600 of 0.05. At an OD600 of 1.0 and 2.0, a total of an OD600 of 2.0 was pelleted at 4000 RPM for 5 min. Supernatant was discarded and pellets were resuspended in 1 mL QIAzol containing lysis beads. Samples were lysed by bead-beating at maximum RPM for 1 min with chilling for 5 min. Bead-beating was repeated twice. Chloroform was added, shaken vigorously, and centrifuged for 15 min at 12,000 x g at 4˚C. 450 µL of the upper phase was combined with 675 µL of 100% ethanol. RNA was extracted using a RNeasy kit, and RNA was eluted using RNase-free water. rRNA depletion, library generation, and >20 million 150-bp paired-end Illumina HiSeq reads were generated for each sample commercially (Genewiz; Azenta).
identifiers.namespace: GEO
identifiers.id: GSM8838473_r1