design: RNA sequencing libraries were prepared with Illumina-compatible NEBNext Ultra II Directional RNA Library Prep Kit. 200 - 500 ng of Qubit quantified total RNA was used for rRNA removal using the NEBNextrRNA Depletion Kit (BACTERIA). Ribodepletion was performed by following manufacturers instructions. Briefly, hybridization of RNA to biotinylated rRNA-specific probes (DNA-based) was carried out at 95C 2 minutes, followed by ramping down the complexes to 22C, and then followed by RNAse H and DNase I digestion. Further the samples were subjected to 1.8X cleanup and eluted in 7ul NFW. The rRNA-free and cleaned-up RNA was quantified on Qubit fluorimeter. The enriched RNA was subjected to fragmentation , first strand synthesis followed by second strand synthesis. The double stranded cDNA was purified with NEBNext purification beads. The cDNA was end-repaired, adenylated and ligated to Illumina multiplex barcode adapters as per NEBNext Ultra II Directional RNA Library Prep protocol followed by second strand excision using USER enzyme at 37 C for 15mins. Adapter ligated cDNA was purified using NEBNext purification beads and was subjected to 12 cycles for Indexing- (98C for 30 sec, cycling (98C for 10sec, 65C for 75sec) and 65C for 5min) to enrich the adapter-ligated fragments. Final PCR product (sequencing library) was purified with NEBNext purification beads, followed by library quality control check.