title: GSM9240606: RBMXL3 IP, rep2; Homo sapiens; RIP-Seq
library_construction_protocol: Two biological replicates of 20 million cells (TCam-2 transfected with RBMXL3_FLAG and GFP_FLAG) were used in eCLIP-seq. Briefly, 48 hours after transfection, TCam-2 cells grown on culture dishes were crosslinked at 254 nm UV with an energy setting of 400 mJ/cm² (Analytik Jena, CL-3000). The cells were then harvested in ice-cold DPBS, and the pellets were flash-frozen in liquid nitrogen and stored at -80°C before the IP reaction with an anti-FLAG antibody, performed by Eclipse BioInnovations (USA). During IP reaction Dynabeads M-280 Sheep Anti-Mouse IgG (Thermo Fisher cat. 11201D) and antibody Anti-FLAG M2 (Sigma Aldrich cat. F1804-1MG) were used. Libraries were constructed using the eCLIP protocol as previously described (van Nostrand et al, 2016).