GSM8214923: T47D, Normoxia_3, input; Homo sapiens; RIP-Seq
Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) following the manufacturer's procedure. The total RNA quality and quantity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number >7.0. Following purification with poly-T oligo attached magnetic beads (Invitrogen), the poly(A) mRNA fractions is fragmented mRNA was subjected to incubated for 2 h at 4℃ with m6A-specific antibody (No. 202003, Synaptic Systems, Germany). Eluted m6A-containing fragments (IP) and untreated input control fragments are converted to final cDNA library in accordance with a strand-specific library preparation by dUTP method. Then we performed the paired-end 2×150 bp sequencing on an Illumina Novaseq™ 6000 platform at the LC-BIO Bio-tech ltd (Hangzhou, China) following the vendor's recommended protocol. m6A-seq