100 ng of total RNA material was used. mRNA was first enriched using oligo(dT) beads and was fragmented randomly by adding fragmentation buffer. Then, cDNA was synthesized using random primers, after which a second-strand synthesis buffer, dNTPs, RNase H and DNA polymerase I were added to initiate the second-strand synthesis. Following end repair and sequencing adaptor ligation, the double-stranded cDNA library was completed through size selection and PCR enrichment. The quality of the final libraries was determined on the Agilent Bioanalyzer with the High Sensitivity DNA kit. The concentration of the libraries was estimated by performing qPCR using a dilution of PhiX Control v3 as standard and samples were pooled in equimolar quantities. Finally, high coverage sequencing was performed on the Novaseq 6000 system with the S4 flowcell and SE50 configuration.