GSM8703113: HEK293 cells transfected with empty vector (NC) for 48 hours, sample 2; Homo sapiens; RNA-Seq
For RNA-seq, mRNA was enriched by polyA(+) selection. Total RNA was qualitatively examined using Nanodrop and Agilent 2000. The construction of long-read sequencing for cDNA libraries was performed using the Oxford Nanopore Technologies PCR-amplified cDNA kit (SQK-PCS109) at Benagene Company (Wuhan, China). Specifically, full-length cDNA libraries were constructed from poly(A)+ mRNA using the cDNA-PCR Sequencing kit. Subsequently, the cDNA was PCR-amplified for 13–14 cycles with barcoded adapters from the Oxford Nanopore PCR Barcoding kit (SQK-PBK004). Then the cell RNA was sequenced by a PromethION platform (P48-seq, the third generation sequencing). the 1D sequencing adapter was ligated to the cDNA before loading onto an R9.4.1 FLO PRO002 flow cell for sequencing on a PromethION platform. The raw data of Nanopore sequencing downstream data are quality controlled, and the obtained sequences are filtered for low-quality data to obtain effective data. Based on the filtered data, the high-quality full-length sequences are obtained. The full-length sequences are compared with the reference genome sequences for visualization.