Full-length transcriptome sequencing of serum: Homo sapiens
We prepared 1 g total RNA for building cDNA libraries using cDNA-PCR Sequencing Kit (SQKLSK110+EXP-PCB096) provided by ONT. Full-length cDNA was richened in template switching activity by reverse transcriptase, and defined PCR adapters were added to both ends of the first strand of cDNA. Then, cDNA was subjected to PCR for 14 circles using LongAmp Tag (NEB), followed by the PCR products connected to ONT adaptor ligation with T4 DNA ligase (NEB). XP reagent beads were applied to DNA purification based on ONT instructions. Finally, the cDNA libraries were loaded on FLO-MIN109 flow cells and sequenced on PromethION platform (Biomarker Technology Company, Beijing, China).