title: GSM8760285: lead acetate-exposed mice, A_3_(Input); Mus musculus; RIP-Seq
library_name: GSM8760285
library_construction_protocol: Total RNA was isolated from cells using the Magzol Reagent (Magen, China)according to the manufacturer's protocol, The quantity and integrity of RNA yield was assessed by using the K5500(Beijing Kaiao, China) and the Agilent 2200 TapeStation (Agilent Technologies, USA) separately. m6A antibody immunoprecipitation RNA was quality control with Qubit(Thermo Fisher Scientifc,USA) and Agilent 2200 TapeStation (Agilent Technologies, USA). mRNA was enriched from total RNA using the DynabeadsTM mRNA Purification Kit (Invitrogen) and then subjected to immunoprecipitation with the GenSeqTM m6A-IP Kit (GenSeq, China). RNA was randomly fragmented into 100–200 nt fragments using RNA fragmentation reagents, and protein A/G beads were coupled to the m6A antibody by rotating at room temperature for 1 h. The RNA fragments were incubated with the bead-linked antibodies and rotated at 4 °C for 4 h to obtain immunoprecipitated RNA fragments. Both the input and m6A IP samples were prepared for the next-generation sequencing.RNA libraries for IP and input samples were then generated with the NEBNext Ultra II Directional RNA Library Prep Kit.