title: Bulk RNA-Sequencing of defined brain regions of mice with acute DSS-induced colitis
design: Bulk RNA-Sequencing of defined brain regions of mice with acute DSS-induced colitis
library_construction_protocol: Male C57BL/6J mice were subjected to 3% DSS dissolved in drinking water for 7 days and sacrificed on day 8 RNA isolation was performed using the RNeasy Mini kit (QIAGEN) following the manufacturerâ€™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.