Pacbio Sequel RNA-Seq of Oreochromis niloticus:healthy Oreochromis niloticus
RNAs from the head kidney, spleen, heart, muscle and liver were mixed equally based on their concentrations and used for PacBio SMRT library generation. The first-strand cDNAs were generated using a SMARTer PCR cDNA Synthesis Kit (Clontech, CA, USA) and the full-length cDNA fragments were selected using BluePippin. Next, PCR was performed for amplification, end-repair and to affix SMRT dumbbell connectors. The samples were then exonuclease digested. Finally, the sequencing library was obtained following a secondary screening using BluePippin (Sage Science, Beverly, MA, USA), with three Iso-Seq libraries (1-2kb, 2-3kb, and 3-10kb) constructed and sequenced with a PacBio system.