Illumina NextSeq500 sequencing and analysis of leaf transcriptomes of Terminalia arjuna
RNA sequencing library was prepared with Illumina-compatible SureSelect Strand-Specific RNA Library Prep kit (Agilent, Santa Clara, CA, U.S.A.) at the Genotypic Technology Pvt. Ltd., Bangalore, India. Briefly, mRNA enrichment was performed from 1 ľg of Qubit quantified total RNA. The enriched mRNA was fragmented for 2 minutes at 94oC in the presence of divalent cations. First strand cDNA was synthesized in the presence of Actinomycin D (Gibco, life technologies, Carlsbad, CA, U.S.A.) and subsequently the single stranded cDNA was purified using HighPrep magnetic beads (Magbio Genomics Inc, USA). Purified single stranded cDNA was used for second strand synthesis followed by end repair using Second Strand Synthesis + End Repair mix. The end repaired cDNA was purified using HighPrep magnetic beads. Adapters were ligated to the cDNA molecules after 3 Adenylation.