GSM6322839: Flag_Cpeb1b_replicate_2; Danio rerio; RIP-Seq
Flag-cpeb1b mRNA injected embryos were collected at 16 hpf and lysed in NETN lysis buffer (150 mM NaCl, 0.5% NP-40, 50 mM Tris-HCl, pH 7.4, RNase inhibitor, Roche cocktail protease inhibitor). The lysate was incubated with anti-FLAG M2 Magnetic Beads (Merck) for 4 h at 4 °C. Then, the beads were washed five times with the lysis buffer and five times with TBS buffer. For RIP-seq assay, the bead-bound RNA was extracted and then fragmented by RNA Fragmentation Reagent (Ambion). The fragmented RNA was subjected to library construction using the SMARTer smRNA-Seq kit (Clontech).