title: RNA-Seq of  mus musculus:adult male testis
design: The total RNAs were isolated from the testis tissues of mice using RNeasy Plus Mini Kit (Qiagen, Hilden, Germany) following the manufacturer's procedure. The RNA integrity was assessed on an Agilent 2100 Bioanalyzer Lab-on-Chip system (Agilent Technologies, Palo Alto, CA, USA). Approximately 1 µg of the total RNA was used to deplete ribosomal RNA according to the manuscript of NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (New England Biolabs, Massachusetts, USA). The rRNA-depleted RNAs were further incubated at 37 °C for 10 minutes with one unit of 20U/μl RNase R (Epicentre, Madison, WI) to digest linear RNAs. The remaining RNAs were used as templates for the construction of cDNA libraries in accordance with the protocol for the NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (New England Biolabs, Massachusetts, USA). The clustering of samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia, San Diego, USA) according to the manufacturer's instructions. And the paired-end sequencing was performed on the Illumina Hiseq1500 platform.