Illumina HiSeq 2000 paired end sequencing; Pseudomonas aeruginosa treated with UV against untreated controls
Pseudomonas aeruginosa treated with UV against untreated controls
Sample 1_p
100 µL suspension of P. aeruginosa was inoculated in the LB medium and cultured overnight at 37°C, with shaking at 150 rpm. Bacteria were harvested by centrifugation at 8,000 rpm for 10 min and then washed thrice with phosphate-buffered saline (PBS, pH 7.2), and suspended in 40 mL PBS (the final concentration of P. aeruginosa was ~109 CFU/mL). For UV irradiation, each 40 mL sample was placed onto a 90-mm-diameter petri dish. A low-pressure (LP) collimated beam apparatus containing an LP (120 W, 30% UVC, TL 120W/01, Philips, Holland) mercury UV lamp was used to perform the UV exposure process. The exposed samples at a UV dosage of 11 mJ/cm2 were preserved for microbial analysis. Bacterial RNA was extracted using the Total RNA Isolation Kit (EZ-10 Spin Column, BBI Life Sciences) according to the manufacturer's instructions. RNA samples that met purity standards (A260/ A280, 1.8e2.0) were submitted to the Beijing Genomic Institute (China) for strand specific cDNA library construction.