title: GSM4504485: irCLIP-Seq data for CENPC in hela cells replicate2; Homo sapiens; RIP-Seq
library_construction_protocol: irCLIP-seq was performed as previously described. Briefly, the 2× 108 isolated B cells were irradiated at 400 mJ/cm2 with 254 nm UV light. The cross-linked cells were lysed and 15 μg of anti-CENP-C antibody was applied to pull down specific protein-RNA complexes. After micrococcal nuclease treatment, 3′ RNA linker ligation and 5′ end 32P labeling, the immunoprecipitated complexes were fractionated on a 4–12% NuPAGE Bis-Tris gel and transferred to a nitrocellulose membrane. The autoradiographed CENP-C-specific smear bands were cut and treated with proteinase K (Takara, Catalog # 9034) prior to the extraction of respective RNA by phenol and chloroform. Strictly follow protocols from NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®.
attributes.tag: GEO Accession
attributes.value: GSM4504485