GSM7719905: Rat kidney [148-2IP]; Rattus norvegicus; RIP-Seq
RNA immunoprecipitation was performed using RIP Kit (Bersinbio, Guangzhou). The kidney lysates from rats were incubated with 5 μg Argonaute-2 (Ago2) antibody (ab186733, Abcam) or a control anti-immunoglobulin G (IgG) antibody (Abcam) coated beads at 4℃ overnight under rotation. The immunoprecipitates were eluted from the beads with elution buffer and the RNAs were purified. Then, purified RNAs were reverse-transcribed into complementary DNA (cDNA). The libraries were prepared according to the manufacturer's instructions and sequenced on an Illumina HiSeqTM 2500 platform by Guangzhou RiboBio Co., Ltd. Antibody immunoprecipitation RNA was quality control with QubitTM(Thermo Fisher Scientifc,USA) and Agilent 2200 TapeStation (Agilent Technologies, USA). Then, 100ng RNA was used for library building following NEBNext® Ultra™ RNA Library Prep Kit protocol for Illumina (NEB, USA). The final library product was assessed with Agilent 2200 TapeStation andQubit®2.0(Life Technologies, USA)