GSM8887888: BXPC3, rep2; Homo sapiens; RNA-Seq
Total RNA was extracted from the samples using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs), following the manufacturer's protocol to selectively enrich for poly(A) RNA. For long-read RNA sequencing, cDNA synthesis was performed using the strand-switching protocol provided by Oxford Nanopore Technologies (ONT). In this protocol, the first-strand synthesis incorporates a unique sequence at the 3' end of the cDNA to enable strand-specific information. Full-length cDNA libraries were then prepared from the poly(A)-selected mRNA using the ONT cDNA-PCR Sequencing Kit (SQK-PCS109). The cDNA was amplified through PCR for 13-14 cycles, using specific barcoded adapters from the Oxford Nanopore PCR Barcoding Kit (SQKPBK004) to facilitate sample multiplexing. After PCR amplification, a 1D sequencing adapter was ligated to the cDNA to prepare the samples for sequencing.