EIF4A3_HA_eif4a3_HA_rep1 (CRX1754724; CRA026448)
Seedlings were crosslinked by 1% formaldehyde and the target RNA-protein complexes were immunoprecipitated by incubating extracted chromatins with Pierce anti-HA magnetic beads (Thermo Scientific, Waltham, MA, USA). After reversal crosslink, DNA removement by TURBO DNase (Thermo Scientific, Waltham, MA, USA) and RNA purification using Tris-Phenol and chloroform, the purified RNA was reverse-transcribed by using random primers and SuperScript III reverse transcriptase (Thermo Scientific, Waltham, MA, USA) with Sorbitol/Trehalose. The single-strand RNA on RNA/cDNA hybrid molecules was digested with RNase ONE Ribonuclease (Promega, Madison, WI, USA) and Ribonuclease H (Thermo Scientific, Waltham, MA, USA). The sequencing libraries were constructed by VAHTS ssDNA Library Prep Kit for Illumina (Vazyme, Nanjing, China) and sequenced on an Illumina NovaSeq 6000 platform.