RNA-seq of SD rats: 8 weeks male skin
In this process, the total RNA was extracted using the Trizol kit. RNA quality was assessed on an Agilent 2100 Bioanalyze and detected using RNase-free agarose gel electrophoresis. After total RNA extraction, eukaryotic mRNA was enriched with Oligo(dT) beads. The enriched mRNA was then fragmented into short fragments using fragmentation buffer and reverse transcribed into cDNA for Illumina. Ligation reactions were purified with AMPure XP Beads (1.0X) and amplified by polymerase chain reaction (PCR). Then, the resulting cDNA libraries were sequenced.The bioinformatics analysis included: 1)Filtering of clean reads by fastp (version 0.18.0) ; 2)Alignment with Ribosome RNA (rRNA) by short reads alignment tool Bowtie2 (version 2.2.8) ;3)Alignment with Reference Genome. Paired-end clean reads were mapped to the reference genome using HISAT2 2.1.0 and other parameters set as a default. 4)Quantification of Gene Abundance. The mapped reads of each sample were assembled by using StringTie v1.3.1 in a reference-based approach .