title: GSM2399854: Clr3_PAR-CLIP_1; Schizosaccharomyces pombe; RIP-Seq
library_construction_protocol: Cells were UV-irradiated (12 J/cm2 at 365 nm) and lysed by bead beating and the lysate was sonicated. Immunoprecipitation was performed on a rotating wheel for 2 h at 4 °C with rabbit IgG-conjugated Protein G magnetic Dynabeads (Invitrogen). Crosslinked RNA was partially digested with RNase T1 and prepared for cDNA library preparation. Data acquisition were performed as described by Schulz and Schwalb et al. (Cell, 2013) with minor modifications. Following adapter ligation, RNA was recovered by Proteinase K digestion for 2 h at 55 °C, and subsequent acidic phenol/chloroform extraction. Reverse transcription was done for 1 hr at 44 °C using SuperScript III RTase (Invitrogen). PCR amplification was performed using the NEXTflex barcode primer kit (Bioo Scientific).
identifiers.namespace: GEO
identifiers.id: GSM2399854