GSM5597020: HeLa-WT-3-INP; Homo sapiens; RIP-Seq
Ribo depleted Total RNA RNA immunoprecipitation was performed using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Merck) following the manufacturer's instructions. Briefly, HeLa cells were transfected with pFLAG-CMV2-DNMT1 plasmid using LipofectamineTM 2000 (Invitrogen) according to the manufacturer's protocol. After 24 h, cells were lysed and the protein concentration was quantified using DCTM Protein Assay Kit (Bio-Rad).  Then, 1.5 mg of proteins were engaged in the IP with 5 µg of anti-FLAG (Sigma) or mouse IgG (control). After washes and proteinase K digestion, immunoprecipitated RNA and total RNA (Input) were subjected to downstream library preparation. The library preparation was performed using TruSeq Stranded Total RNA with Ribo-Zero Gold following the manufacturer's instructions.