RNA-Seq analysis of USB1: USB1 overexpression in Kasumi-1 cells
Kasumi-1 cells with USB1 overexpression and corresponding control cells were collected for RNA extraction. Total RNA was isolated using TRIzol reagent according to the manufacturers instructions. RNA integrity and concentration were assessed prior to library preparation. mRNA was enriched using poly(A) selection and subsequently fragmented into short fragments. First-strand cDNA was synthesized using random hexamer primers, followed by second-strand cDNA synthesis. The cDNA fragments were end-repaired, A-tailed, and ligated to sequencing adapters. The libraries were amplified by PCR and purified. Library quality was evaluated before sequencing on an Illumina platform.