title: GSM5911570: H1 NAT10-KD rep2 input; Homo sapiens; RIP-Seq
library_construction_protocol: Cells from each group were collected and total RNA was harvested with Trizol reagent. Remove the rRNAs from the total RNA by using the Ribo-Zero rRNA Removal Kits (Illumina, USA). The RNA was randomly fragmented into fragments of about 200 nt.The Protein A/G magnetic beads were incubated with the ac4C antibody by rotation for 1 h at room temperature to bind the antibody to the magnetic beads.The RNA fragment was then incubated with a 4°rotation of the antibody bound to the magnetic beads for 4 h to bind the RNA to the antibody.The RNA/antibody complex was eluted from the magnetic beads by Protease K, and the RNA was extracted by the phenol: chloroform method. RNA sequencing libraries were constructed using the NEBNext® Ultra II Directional RNA Library Prep Kit. The constructed libraries were quality-controlled with Agilent 2100 bioanalyzer and then sequenced on Illumina's NovaSeq sequencer for high-throughput sequencing.