Transcriptome Analysis of 18 Tissue Samples

Total RNA was extracted from 18 tissues (6 samples in 3 repeats) using the TaKaRa RNA extraction kit, and cDNA libraries were prepared. The mRNA was obtained using Oligotex mRNA Midi Kit (Qiagen, Germany). The RNA quality was assessed using a Nano-Drop spectrophotometer (Thermo Fisher Scientific, United States), followed by cDNA library preparation using the Illumina protocol (Ahmad et al., 2021a (link)). The library products were evaluated with the Agilent 2200 TapeStation and Qubit^®2.0 (Life Technologies, United States). The products were diluted to 10 pM for the in situ generation of clusters on HiSeq2500 pair-end flow cells, followed by pair-end sequencing (2 × 100 bp). Finally, transcriptome de novo was performed with the Trinity program using default parameters (Grabherr et al., 2011 (link)). Gene expressions were quantified using fragments per kilobase per transcript per million mapped reads (FPKM).