Rota 3
50ng of total RNA was taken for fragmentation and priming. Fragmented and primed RNA was further subjected to first strand synthesis followed by second strand synthesis. The double stranded cDNA was purified using JetSeq Beads (Bioline, Cat # BIO-68031). Purified 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