title: Illumina NovaSeq 6000 paired end sequencing; Transcriptional profiling of murine distal colon from C57BL/6 (WT) and Stat2KO mice during acute DSS colitis.
design: Transcriptional profiling of murine distal colon from C57BL/6 (WT) and Stat2KO mice during acute DSS colitis.
library_construction_protocol: Mice were sacrificed and its colon extracted post-mortem RNA was isolated using the PeqGold Total RNA Kit following manufacturers' instructions (Peqlab/VWR). 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.