BG4 RIP-seq using mouse forebrain tissues

To identify G4-enriched mRNAs expressed in the mouse brain, RNA immunoprecipitation by BG4 antibody (G4-specific antibody) was performed. Overall design: Mouse forebrain tissues were lysed and precleared using the Protein G HP SpinTrap Kit (GE healthcare). The precleared samples were incubated at 4°C overnight in a Protein G HP SpinTrap column that had been treated with His-tagged BG4 (or normal rabbit IgG for control) and anti-6×His. The resultant solution was washed three times and eluted with elution buffer. The BG4-bound RNA fraction was purified using the RNeasy Mini Kit (Qiagen) and subjected to RNA-sequencing library preparation.