Bulk RNA-Sequencing of brain hemispheres of mice with colitis caused by Citrobacter rodentium infection
Bulk RNA-Sequencing of brain hemispheres of mice with colitis caused by Citrobacter rodentium infection
On day 8  post infection, mice were sacrificed, transcardially perfused with PBS and right brain hemispheres collected After 8 hours of fasting, male C57BL/6J mice were infected with 109 colony-forming units of C. rodentium strain ICC169 in PBS by oral gavage. Control mice received 200µL PBS orally. RNA isolation was performed using the RNeasy Mini kit (QIAGEN) following the manufacturer's instructions. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.