GSM4877083: Cortex Trisomy rep3 Input; Homo sapiens; OTHER
Total RNA was extracted with Trizol (Invitrogen). Polyadenylated RNA was isolated by Sigma GenElute mRNA miniprep kit. Purified mRNA was randomly fragmented via RNA fragmentation Reagents (Millipore). m6A antibody (Synaptic Systems) was incubated with Magnetic Beads A/G Blend (Millipore). Afterwards, fragmented mRNA was added to m6A antibody-beads mixture. The eluted RNA was extracted with Acid Phenol followed by standard ethanol precipitation. The TruSeq Stranded mRNA Sample Prep Kit (Illumina) was used for library preparation both on m6A IP and input samples, and then the library was deeply sequenced on an Illumina HiSeq platform.