RNA-seq of mus
The library was prepared for Illumina sequencing using the TruSeq Stranded mRNA Library Prep Kit. Total RNA was extracted from cell samples using the TRIzol reagent and RNA quality was assessed using the Agilent 2100 Bioanalyzer. mRNA was enriched using oligo(dT) magnetic beads and fragmented using divalent cations. First-strand cDNA was synthesized using random hexamers and SuperScript II Reverse Transcriptase. Second-strand cDNA synthesis and end repair were performed using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina. Adapters were ligated using the TruSeq RNA UD Adapters and the library was amplified using PCR. Library size was confirmed to be approximately 300bp using the Agilent 2100 Bioanalyzer, and library concentration was quantified using qPCR. The final library was sequenced on an Illumina platform using a 2x150bp paired-end sequencing strategy