RNA-seq Analysis of Mice Transcriptomes

Three biological replicates (three mice) from each age group were used in the RNA-seq analysis. Total RNA extracted above was subjected to rRNA depletion by Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina, San Diego, CA, United States). Strand-specific RNA libraries were prepared using TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) without poly-A selection. All assays were performed according to the manufacturer’s instructions. Sequencing was carried out at the Illumina Nova6000 plate High Output Model (Illumina, San Diego, CA, United States) (Hrdlickova et al., 2017 (link)), in a 2 × 150 bp paired-end configuration, with a total of more than 2,666 M reads per lane (at least 40 M reads per sample). All sequencing data are available in NCBI database (accession number: SRP271007).