GSM5525951: Ctrl-Input; Mus musculus; RIP-Seq
RNA in cells was extracted by Trizol, m6A on mRNA was specifically identified by m6A antibody, and methylated RNA fragments were enriched by RNA immunoprecipitation method The mRNA m6A was sequenced by MeRIP-seq at Novogene (Beijing, China). Briefly, a total of 300 µg RNAs were extracted from the Min6 cells. The integrity and concentration of extracted RNAs were detected using an Agilent 2100 bioanalyzer (Agilent) and simpliNano spectrophotometer (GE Healthcare), respectively. Fragmented mRNA (~100 nt) was incubated for 2 hr at 4℃ with anti-m6A polyclonal antibody (Synaptic Systems) in the immunoprecipitation experiment. Then, immunoprecipitated mRNAs or Input was used for library construction with NEBNext ultra RNA library prepare kit for Illumina (New England Biolabs). The library preparations were sequenced on an Illumina Novaseq or Hiseq platform with a paired-end read length of 150 bp according to the standard protocols.