GSM8228768: Trim21_KO_1_UV; Mus musculus; RNA-Seq
spleens  were harvested following final UVB exposure. To generate a single cell suspension, spleens were passed through a 70-μM filter. RNA was isolated using TRI Reagent (Millipore Sigma). Total RNA samples were analyzed for RNA integrity on the 2100 Bioanalyzer using the Agilent RNA 6000 Nano Kit (Agilent Technologies, Santa Clara, CA) and quantified using the Qubit RNA HS Assay Kit (ThermoFisher Scientific, Waltham, MA). Three hundred ng of total RNA Total was ribodepleted using the RiboCop Depletion Kit Human/Mouse/Rat v2 (Lexogen Inc., Greenland, NH).  Stranded RNA-Seq library construction was performed using the xGen Broad-Range RNA Library Prep Kit (Integrated DNA Technologies, Coralville, IA).  Library concentration was measured with a Qubit fluorometer (ThermoFisher Scientific), and library size was evaluated on a 4200 TapeStation (Agilent Technologies). Multiplexed libraries were sequenced on a NovaSeq 6000 (Illumina, San Diego, CA) using 75bp single-end sequencing. On average, approximately 50 million reads were generated from each sample.