GSM8712324: shNC #1 Input; Homo sapiens; RIP-Seq
100μg total RNA was extracted from cells using TRIzol following the manufacturer's protocol (phenol-choloroform extraction). These samples are extracted from human cancer cells. RNA was then purified and treated with DNAse using the Zymo RNA Clean and Concentrator Kit. mRNA was purified using a Dynabeads mRNA DIRECT Kit (Thermo Fisher, # 61012). Samples went through the mRNA extraction protocol twice to eliminate rRNA contamination. 1ug of RNA was then fragmented using the NEB RNA Fragmentation Module to 200nt. 5% of the fragmented mRNA was saved as input. mRNA fragments containing m6A were enriched with an EpiMark N6-Methyladenosine Enrichment Kit (NEB, E1610S) and then extracted using RNA Clean and Concentrator (Zymo Research). 2 biological replicates per each condition were used. Oligo-dT mRNA directional