RNA-seq of Mus musculus C3H/HeJ: uninfected spleen
Spleen tissue samples were collected in RNAlater solution. Total RNA from spleen tissues was isolated by using QIAGEN RNeasy Mini Kit, (QIAGEN, CA, USA) according to manufacturer's instruction. The quality and concentration of the RNA samples were determined by using Agilent Tapestation 2200 system on the RNA screen tape (Agilent Technologies, CA, USA) and Qubit, Broad Range flurometric assay respectively (Thermo Fisher Scientific). All RNA samples were normalized to 80 ng/L for input into the Illumina TruSeq Stranded mRNA LS library preparation kit. Individual libraries were constructed and barcoded according to the manufacturers protocol (Illumina, CA, USA). Sequencing library quality was assessed via Agilent TapeStation 2200 D1000 DNA screen tape (Agilent Technologies) showing a final library size of approximately 270 bp and were quantified with the Qubit High Sensitivity dsDNA assay (Thermo Fisher Scientific). All libraries were normalized and pooled in equimolar concentration for sequencing on an Illumina NextSeq 500 75 cycle High Output kit to generate approximately 400 million 75 base-pair, single-end sequencing reads. All raw data was uploaded to Illumina BaseSpace (basespace.illumina.com) for FASTQ generation and demultiplexing of sequencing reads for their respective samples.