RNA-seqA of Thunnus alalunga: adult male muscle
A total amount of 300ng RNA was used as input material for the RNA sample preparation. Library preparation was performed using Iso-Seq express 2.0 kit (Pacific Biosciences, 103-071-500) and Kinnex full-length RNA kit (Pacific Biosciences, 103-072-000). In brief, first strand cDNA was synthesized by reverse transcription after adding primer with Oligo dT paired with Poly(A).Template switching was performed by adding the other primer paired with TS oligo. Full-length cDNA was obtained by cDNA amplification with barcoded cDNA PCR primers. Perform 8 parallel Kinnex PCR reactions with Kinnex primers to generate DNA fragments containing orientation specific Kinnex segmentation sequences. Pooling of 8 Kinnex PCR products and quantified by Qubit fluorometer (Invitrogen, Q33238). Treat kinnex PCR-amplified cDNA fragments with Kinnex enzyme, ligase, and barcoded Kinnex terminal adapters to assemble cDNA segments into a linear array. Nuclease treatment and cleanup. Library purification using 1X cleanup beads. After quality control test by Agilent Fragment Analyzer System (Agilent Technologies, M5310AA), a SMRTbell library was obtained. The library was sequenced on the PacBio Revio system (Pacific Biosciences, 102-090-600).
SUB16273023
ALB_RNAA