GSM9083061: HeLa, totalRNA, Input, rep1; Homo sapiens; RIP-Seq
Total RNA was isolated from chromatin-associated fraction of SMMC-7721, HepG2, HCT116 and HeLa cells. To isolate mRNA from total RNA, the Dynabeads® mRNA Purification Kit (Invitrogen) was used following the manufacturer's protocols. Non-ribosome RNA was enriched from total RNA by using RiboMinus transcriptome isolation kit (Invitrogen). 1 μL 1:1000 diluted m6A spike-in from EpiMark N6-Methyladenosine Enrichment Kit (NEB #E1610S) was added to 1 μg polyA or non-ribosome RNA, followed by fragmentation according to previously published protocols. m6A-IP was performed using EpiMark N6-Methyladenosine Enrichment Kit following the manufacturer's protocols. Library preparation was performed by using SMARTer Stranded Total RNA-Seq Kit (Takara) v2 following manufacturer's protocols.