GSM5209335: Input (poliA) Rep 1; Homo sapiens; RIP-Seq
HEK-293T cells were transfected with the HIV-2 pROD10-ΔEnv molecular clone. At 24 hpt, total RNA was extracted and 70 microg were treated with kit TURBO DNA-free™️ Kit and then used to obtain polyA RNA using the mRNA isolation kit (Sigma-Aldrich). PolyA RNA was fragmented using the RNA Fragmentation Reagent (Ambion), 500 ng were stored as the input and 2 microg were used for immunoprecipitation of the methylated mRNA (m6A-IP samples) using an anti-m6A antibody (0,5 mg/mL; Synaptic System #202003). cDNA libraries preparations and RNAseq was performed as a service from Genoma Mayor. All the samples were sequenced in an Illumina HiSeq2000 platform with paired-end 150 bp read length. This was obtained as a  RNA Seq/TruSeq Stranded RNA + RiboMinus. meRIP-seq