title: Pam_IR_634 wildtype 5 hrs HIM rep1
design: RNA sequencing was performed as per in vitro culturing with three x 2 mL aliquots of strains in KB to represent initial uninduced expression and HIM for induced expression taken at specific time points, centrifuged at max speed for 5 mins to pellet the cells and the supernatant removed, after which they were frozen in liquid nitrogen and stored at 80 C until extractions. RNA extractions were performed using Trizol and chloroform extraction, followed by Qiagen column cleaning per the manufacturers instructions using the RNeasy kit and elution in 30 L of DEPC. Bacterial DNA was removed post column via Turbo DNase (Invitrogen, USA) treatment and phenol:chloroform:isoamyl alcohol with 1M Tris buffer (Invitrogen, USA) clean up. RNA quality was assessed via nanodrop checking for ratios of >2 for 260:280 and 260:230, concentration was determined using Qubit RNA kit and degradation via analysis of bands on a 1% agarose gel. Pure RNA was then sent to Novogene. Samples underwent QC including Nanodrop, Agilent 5400 and agarose gel electrophoresis. Samples were library prepped using the Prokaryotic mRNA-seq (WOBI) workflow. They were sequenced using paired-end Illumina sequencing with Novogene (NovaSeq X Plus Series (PE150) (2 G raw data per sample).
library_name: Pam_IR_634_WT_5h_R1
identifiers.namespace: SUB15972350
identifiers.id: Pam_IR_634_WT_5h_R1