GSM4903826: Dermal fibroblasts PID21_NS; Homo sapiens; RIP-Seq
Day 14 and day 21 cells were harvested for HITS-CLIP following the published protocol (Moore et al., 2014) with modifications. Briefly, cross-linked cells were lysed and treated with RQ1 DNase (Promega, M6101) and RNaseA (Thermo). Complex containing AGO-miRNA-mRNA were immunoprecipitated overnight at 4C with pan-AGO antibody. The immunoprecipitated complex was radio-labelled and extracted after running on NuPAGE gel (Thermo). RNA from the 130kDa band was extracted for sequencing using TruSeq Small RNA Library Preparation Kits (San Diego, CA). TruSeq Small RNA Library Preparation Kits (San Diego, CA).