title: GSM8440400: Control1.IP; Mus musculus; RIP-Seq
library_construction_protocol: Control organoids  were collected in ice-cold PBS solution and then incubated in RIP buffer (150 mM KCl, 0.5 mM DTT, 25 mM Tris, pH 7.4, 1 mM PMSF, 0.5% NP40 and protease Inhibitor) after centrifugation. Organoids in RIP buffer were then rocked on a nutator at 4°C for 30 min, then cell membrane and debris were discarded by centrifugation at 4°C for 15 min. The collected supernatant was then added to either antibody anti-IgG or anti-RBM25 and incubated at 4°C overnight. Protein A/G Dynabeads was added and incubated at room temperature for 2 h. Dynabeads were then washed 5 times each for 10 min with washing buffer (1 M NaCl, 1% NP-40, 1 mM EDTA, 50 mM Tris, pH 7.4, 0.1% SDS, and 0.5% sodium deoxycholate) and then subjected to RNA extraction. RNAs were extracted from the Dynabeads using Trizol. Extracted RNAs from RIP samples and then underwent library construction on the Illumina NovaSeq 6000 sequence.