title: Illumina HiSeq 3000 paired end sequencing; RNAseq analysis of Campylobacter jejuni exposed to 405nm light
design: RNAseq analysis of Campylobacter jejuni exposed to 405nm light
library_construction_protocol: 1.5 ml of culture was collected and centrifuged (12,000 g, 2 min, 4oC). The resulting supernatant was removed, and cell pellets were immediately frozen by submerging in liquid nitrogen for 45 seconds C. jejuni samples were grown from an overnight starter culture. Cells were harvested by centrifugation and resuspended in 50 ml BTS to an OD600 0.1 and added to 6 well plates (Greiner), 5 ml per well. After growth to mid-log (OD600 =0.6) phase, the T0 timepoint was taken and cells were exposed to 405 nm light with samples taken after 15 and 30 min light exposure (18 mW cm2). Cells were treated with high intensity 405 nm light produced by a Thorlabsâ€™ M405L3 mounted fibre-coupled LED with a nominal wavelength of 405 nm and a bandwidth of ~20 nm at full half width maximum. Power density (J cm-2) was measured using a Thorlabs 5310C thermal power sensor 200 mm away from the light source at the position the sample cuvette is mounted. Cells were exposed to light at mid log phase for the amount of time indicated in the sample list RNA isolation was performed by GENEWIZ rRNA depletion and transcript library preparations were performed by GENEWIZ