title: RNA-seq of Mus musculus(C57):spleen-Spen-2
design: To isolate the total RNA from the tissues, we used the RNeasy Plus Universal Mini Kit (Qiagen, Germantown, MA, USA, Cat. #73,404). RNA quality was assessed by 1% agarose gel electrophoresis and a NanoPhotometer spectrophotometer (Implen, Westlake Village, CA, USA). RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). RNA-Seq libraries were generated with the NEBNext Ultra Directional RNA Library Prep Kits (NEB, USA) according to the manufacturers protocol. After clusters generation on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia), the libraries were sequenced on an Illumina Novaseq platform (Novogene, Tianjin, China) with paired-end 150 bp.