Transcriptome of Amphiprion percula infected with Cryptocaryon irritans
The mRNA was enriched from the total RNA using Poly-T attached magnetic beads, followed by enzymatic fragmentation, 1st strand cDNA conversion using SuperScript II and Act-D mix to facilitate RNA dependent synthesis. The 1st strand cDNA was then synthesized to 2nd strand using the 2nd strand mix. The double strand cDNA was then purified using the AMPure XP system (Beckman Coulter, Beverly, MA, USA) followed by A-tailing, adapter ligation and then enriched by limited number of PCR cycles. The PCR products were purified by the AMPure XP system and the quality of PCR enriched libraries was assessed on 4200 Tape Station system (Agilent Technologies) using high sensitivity D1000 Screen tape as per manufacturer instructions.
Fish gill transcriptome