GSM6742526: HBV-KMT2B integrated HCC Iso-seq; Homo sapiens; OTHER
Total RNA was extracted by RNeasy Mini (QIAGEN, Venlo, Netherlands). Firstly, the first strand cDNA was synthesized from the polyA+ tail by NEBNext's reverse transcriptase. Then the enzyme's terminal transferase activity added a few extra nucleotides to the 3' end of the cDNA. Subsequently template switching, extension and amplification were performed to generate double-stranded cDNA. The cDNA was ligated with universal hairpin adapters. After purification with magnetic beads , sequencing primer was annealed to the SMRTbell templates, followed by binding of the sequence polymerase to the annealed templates.