Full-length transcriptome from Acipenser schrenckii
To construct the library for PacBio sequencing, qualified RNA from seven tissues, including the three testes and four ovaries, were mixed in equal amounts. The mixed RNA sample was reverse-transcribed for mRNA using the SMARTerTM PCR cDNA Synthesis Kit. The PCR amplification was performed using the KAPA HiFi HotStart PCR Kit.Then, the PCR product for the SMRTbell library was constructed using the SMRTbell template pre kit. The concentration of the SMRTbell library were measured using Qubit 3.0 fluorometer with QubitTM 1X dsDNA HS Assay kit (Invitrogen, Carlsbad, USA). The quantified criteria of the library quality were concentration > 10 ng/l with dispersive but continuous distribution in the range of 1-10k. A total of 2.5 ng of the library for each SMRT cell was sequenced using binding kit 2.1 on the PacBio Sequel platform with 20 hours of movie times.