title: GSM8484026: Cre_IP_insulin, adiocytes; Mus musculus; RIP-Seq
library_construction_protocol: Cells were harvested with cell scraper and transferred to an Eppendorf tube. Cell lysate was further homogenized by passage through a 25G syringe needle 10 times. The lysate was sonicated with a cup horn (Q700, QSONICA) for 3 cycles (30s ON/OFF, 50 Amp) to break the nuclei (microscopic examination) and to shred the RNA. Supernatant was collected by centrifugation at 13,000 RPM at 4oC. The immunoprecipitated RNA was quantified with the Qubit RNA HS assay kit (Thermo Scientific) before rRNA Removal with NEBNext® rRNA Depletion Kit v2. The entire rRNA-depleted RNA was used for sequencing library construction.