RNA-seq of Andrographis paniculata

Total RNA with RIN value above 7.0 was processed further for cDNA library preparation. The cDNA library was prepared using Ion total RNA seq kit v2 (Ion Proton™). mRNAs from the total RNA were enriched by depleting rRNA using Ribominus plant kit (Invitrogen). The rRNA depleted total RNA was further fragmented enzymatically using RNase III enzyme to obtain fragments of 200bp and purified using Agencourt Ampure Beads (Beckman Coulter, Life Sciences). Subsequently, fragmented rRNA depleted total RNA served as templates for reverse transcription and ligated with adaptors followed by PCR amplification. Finally, the library was enriched on Ion OneTouch2 and Ion OneTouch2 ES system (Thermo Fisher Scientific) using Dynabeads™ MyOne™ Streptavidin C1 (Invitrogen). Following this, the enriched library were loaded on Ion PI™ Chip Kit v3 (Ion Torrent) and sequenced on Ion Proton™ System (Ion Torrent).