GSM8947569: RIP-seq-HEK293T-Ctrlrep2; Homo sapiens; RIP-Seq
Total RNA was extracted from HEK293T or other cell lines using TRIzol™ reagent (Thermo Scientific, 15596018) following the manufacturer's instructions. Total RNA was extracted from HEK293T and HuCCT1 cells using TRIzol™ reagent according to the manufacturer's instructions. RNA-seq library preparation was conducted using the Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit, following the manufacturer's protocol. This process included ribosomal RNA depletion to enrich the library for mRNA and pre-mRNAs. After rRNA depletion, the RNA was fragmented and denatured to generate appropriate RNA fragments for cDNA synthesis. First-strand cDNA was synthesized, and an A-tail was subsequently added to the 3' end of the cDNA to facilitate adapter ligation. Illumina sequencing adapters were then ligated to the cDNA fragments, and the adapter-ligated library was amplified via PCR to enrich for the desired fragments. The final RNA-seq library was quantified and normalized to ensure uniform sequencing depth across samples. High-throughput sequencing was performed on the Illumina HiSeq2500 platform.