GSM7610448: SW1990 cell,Mock-1 [RNA-seq]; Homo sapiens; RNA-Seq
RNA was harvested using Trizol (Takara). A total amount of 1μg qualified RNA per sample was used as input material for the library preparation. Total RNA was isolated from cells/tissues using the Magzol Reagent (Magen, China) according to the manufacturer's protocol, The quantity and integrity of RNA yield was assessed by using the K5500 (Beijing Kaiao, China) and the Agilent 2200 TapeStation (Agilent Technologies, USA) separately. Briefly, the mRNA was enriched by OligodT according to instructions of NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB, USA). And then fragmented to approximately 200bp. Subsequently, the RNA fragments were subjected to first strand and second strand cDNA synthesis following by adaptor ligation and enrichment with a low-cycleaccording to in structions of NEBNext® Ultra™ RNA Library Prep Kit for Illumina. The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit (Thermo Fisher Scientific, USA). The libraries were sequenced by Illumina (Illumina, USA) with paired-end 150bp at Ribobio Co. Ltd (Ribobio, China). The clean reads were obtained after removal of reads containing adapter, ploy-N and at low quality from rawdata. HISAT2 was used to align the clean reads to the human reference genome hg38 with default parameters. HTSeq was subsequently employed to convert aligned short reads into read counts for each gene model. Differential expression was assessed by DEseq/DESeq2/edgeR/DEGseq using read counts as input. The Benjamini–Hochberg multiple test correction method was enabled.