cDNA library preparation and sequencing

TruSeq RNA sample preparation Kit v2 (Illumina, San Diego, California, USA) was used To generate paired-end cDNA sequencing libraries. The library was prepared using total RNA (1 μg) by following the manufacturer’s protocol. To enhance the sequencing depth, Oligo-dT covered magnetic beads were used to purify poly-A attached mRNA by eliminating other RNA molecules. The fragmentation of enriched and purified mRNA was subjected using fragmentation buffer at 94 °C temperature in a pre-heated thermal cycler. The fragmented mRNA was reverse transcribed into the first-strand cDNA using random primers. In the presence of DNA polymerase I and RNase H, synthesis of the second-strand cDNA was performed. After end repairing and 3′ adenylation, adapters were ligated to the cDNA ends to make ready for hybridization to a single-read flow cell. Before library preparation, cDNA fragments were purified and enriched with PCR. The quality of the library was examined by DNA-1000 kit using Agilent 2100 Bioanalyzer (Agilent Technologies, California, USA). Each library was sequenced in the MiSeq platform (Illumina, San Diego, California, USA) by MiSeq Reagent Kit v2 (150 × 2 bp). The sequence data generated in present study have been deposited at NCBI in the Short/Sequence Read Archive database under the accession numbers: SRR12791806 and SRR12791807 with Bioproject ID: PRJNA667584.