Test KS10_1a
500 ng of total RNA was taken for mRNA isolation, fragmentation and priming. Fragmented and primed mRNA was further subjected to first strand synthesis followed by second strand synthesis. The double stranded cDNA was purified using JetSeq Beads. Purified cDNA was end-repaired, adenylated and ligated to Illumina adapters as per NEBNext UltraTM 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 JetSeq Beads and was subjected to 11 cycles for Indexing-(98 C for 30 sec, cycling and enrich the adapter-ligated fragments. Final PCR products were purified with JetSeq Beads, followed by library quality control check. Illumina-compatible sequencing libraries were quantified by Qubit fluorometer and fragment size distribution was analyzed on Agilent 2200 Tapestation.