P12_Tumor_PacBio_IsoSeq
Tissue samples were lysed with RiboEx solution, and pure RNA was recovered via glass-fiber membrane column purification. RNA concentration and purity were evaluated using a NanoDrop spectrophotometer and a Qubit 4 Fluorometer with the RNA HS Assay Kit (Thermo Fisher Scientific, MA, USA), while RNA integrity was confirmed on an Agilent 2100 Bioanalyzer using the RNA 6000 Nano Kit (Agilent Technologies, CA, USA). For long-read transcriptome sequencing, Kinnex Iso-Seq libraries were prepared using the Iso-Seq Express 2.0 Kit (Pacific Biosciences, CA, USA). Briefly, full-length cDNA was synthesized with specific handles, and 5 barcodes were added during cDNA amplification. To optimize sequencing throughput, Kinnex adapter ligation was performed to concatenate cDNA arrays. Final library quantification and quality assessment were performed using the Qubit dsDNA HS Assay Kit and the Agilent 2100 Bioanalyzer with the DNA 12000 Kit, respectively. Libraries were sequenced on a PacBio Revio system (Pacific Biosciences, CA, USA).
SUB16217207
P12_Tumor_PacBio_IsoSeq