GSM7124037: WT Run 1 input replicate 2; Homo sapiens; RIP-Seq
Total RNA was collected using Trizol reagent according to the manufacturer's protocol. Following separation of the aqueous and organic phases, the aqueous phase was removed and purified with a Qiagen RNA kit according to the manufacturer's protocol. mRNA was polyA selected through two rounds of selection with oligo dT magnetic beads. Following polyA selection, mRNA was fragmented using ZnCl2 before selection with m6A-specific antibody. 3' adaptors sequencing adaptors were added with RNA ligase and reactions were reverse-transcribed with SuperScript IV. cDNA was circularized with circLigase and libraries were PCR amplified to add Illumina sequencing primers. Libraries were sequenced on a NextSeq 550 instrument.