GSM7898506: K562-wt SF3B1 iCLIP replicate 2; Homo sapiens; RIP-Seq
iCLIP was performed as described in Huppertz et al, Methods, 2014, PMID:24184352. Exponentially growing K562 cells were pelleted, washed once with PBS and 10x10^6 were subjected to UV-cross linking at 400 mJ/cm2 at 254 nm in 6 ml PBS in a 10 cm petri dish on ice. Crosslinked cells were scraped from the dish, collected by centrifugation for 2 min at 500 xg at 4 °C, snap frozen in liquid nitrogen and stored at -80 °C. About 3x10^6 cells were immunoprecipitated using antibody. For iCLIP of K562-SF3B1wt/wt and K562-SF3B1K700E/wt, 10 µg of a monoclonal SF3B1 antibody (clone 16, MBL D221-3) was used. For U2AF2 iCLIP, 10 µg of a monoclonal U2AF2 antibody (Sigma, U4759) was used. iCLIP library preparation was performed as described in Huppertz et al, Methods, 2014, PMID:24184352.