M3 (CRX1754737; CRA026449)
A 500 ng qualified total RNA sample was taken and the volume was adjusted to 9 ul using nuclease free water. Link the reverse transcription primer to carry out reverse transcript. Then mix 5 microL reverse-transcribed RNA sample, 1.5 microL PCR barcode, 18.5 microL nuclease-free water and 25 microL 2x LongAmp Taq Master Mix to start to amplify. AMPure beads were used for purification. 500 ng purified cDNA and Rapid Adapter were used to connect sequencing splices. Finally load the library onto the R9.4 sequencing chip for computer sequencing for 48~72 hours after configuring the library with 24 microL cDNA connected products, 75 microL squencing buffer and 51 microL loading beads. submitted_library_strategy: ISO-Seq submitted_instrument_model: OXFORD_NANOPORE PromethION