TNFa treated B6 WT biological replicate 3
Libraries were prepared using NEB Next Ultra II Directional RNA Library Kit for Illumina according to the manufacturers protocol. Briefly, after mRNA enrichment, the RNA was fragmented at 94oC for 15 min. First strand cDNA was generated, and incorporation of dUTP occurred during synthesis of second strand cDNA to indicate strandedness. Double-stranded cDNA then underwent end repair, A-tailing, adaptor ligation, and incorporation of sample-specific multiplex indices via PCR amplification (11 cycles) according to the manufacturers protocol (New England Biolabs, USA). Size distribution and molarity of amplified cDNA libraries were assessed via the Bioanalyzer High Sensitivity DNA Assay (Agilent Technologies, USA).