GSM5766909: HepG2 D312A rep1 (IP); Homo sapiens; OTHER
GSM5766909
Three biological replicates of 40 million (2✕ 150 mm plates) HepG2 cells of stable overexpression of EGFP, METTL14 (WT), METTL14 (R298P) and METTL14 (D312A) were lysed by Trizol and the total RNA was extracted. The total RNA was treated by DNase I (5 U per 100 µg total RNA) and mRNAs were enriched by NEBNext Poly(A) mRNA magnetic isolation module (NEB E7490L). 8-10 µg of mRNA from each sample was fragmented to the size of ~120-nt by Ambion Fragmentation reagent (Thermo Scientific AM8740) in a proportional volume (10 µl reaction per 1 µg mRNA) at 70 ℃ for 7 min then precipitated in 75% ethanol with 10% volume of sodium acetate (3 M, pH 5.5) and GlycoBlue (1 µ