GSM5548651: METTL3-deficient MDA-MB-231 input rep2; Homo sapiens; OTHER
Total RNA was isolated from samples with TRIzol reagents following the manufacturer's instructions. Poly(A)+ RNAs were purified using Dynabeads mRNA purification kit, followed by fragmentation and immunoprecipitation with anti-m6A antibody in IP buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1% NP-40) for 2 h at 4°C. The m6A-IP mixture was then incubated with Dynabeads protein A for an additional 2 h at 4°C. The bound RNA was eluted by competition with N6-methyladenosine and purified using an RNA cleanup kit.