title: Comparative Transcriptome analysis of chickpea samples: chickpea_elm_2
design: RNA sequencing libraries were prepared with Illumina-compatible NEBNext Ultra II Directional RNA Library Prep Kit. 500 ng of Qubit quantified total RNA was taken for mRNA isolation, fragmentation and priming. Fragmented and primed mRNA was further subjected to first strand cDNA 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 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 HighPrep Beads and was subjected to 10 cycles for Indexing to enrich the adapter-ligated fragments. Final PCR product was purified with HighPrep Beads, followed by library quality control check. Illumina-compatible sequencing library was quantified by Qubit fluorometer and its fragment size distribution was analyzed on Agilent 2200 TapeStation.