title: Transcriptomics of fish gut
design: The qualities and quantities of the isolated RNA samples were checked on 1% denaturing RNA agarose gel and Nanodrop, respectively followed by Agilent Tape station using High Sensitivity RNA screenTape. The RNA-Seq paired end sequencing libraries were prepared using illumina TruSeq Standed mRNA Prep kit. Briefly, mRNA was enriched from the total RNA using Poly-T attached magnetic beads, followed by enzymatic fragmentation, 1st strand cDNA conversion using SuperScript ll and Act-D mix to facilitate RNA dependent synthesis. The 1st strand cDNA was then synthesized to second strand using second strand mix. The dscDNA was then purified using AMPure XP beads followed by A-tailing, adaptor ligation and then enriched by limited no. of PCR cycles.  Quantity and quality check of library was performed on Agilent 4200 tape station using high sensitivity D1000 Screen tape. After obtaining the Qubit concentration for the libraries and the mean peak sizes from Agilent Tape Station profile, the PE ilumina libraries were loaded onto Illumina Platform for cluster generation and sequencing. Paired-End sequencing allows the template fragments to be sequenced in both the forward and reverse directions on Illumina platform.