GSM8415487: WPMY-1, sh61A-1.AlkB-, m1A-MAP-seq; Homo sapiens; OTHER
Total RNA was extracted using Trizol reagent m1A-MAP-Seq service was provided by Cloud-Seq Biotech (Shanghai, China).The MirVana Isolation Kit (ThermoFisher) was used to enrich small RNAs which were less than 200 nt from total RNAs. The m1A-MeRIP reaction was performed using the GenSeq m1A-MeRIP Kit as follows: RNA was incubated with m1A antibody in IPP buffer for 2 h at 4 degrees. The reaction mixture was further immunoprecipitated with protein A magnetic beads (Thermo Fisher) at 4 degrees for 2 h. The bound RNA on the beads was then eluted with a free m1A adenosine analogue. The eluted RNA was further extracted by Trizol reagent (Thermo Fisher). The extracted and purified RNA was divided into two groups and treated respectively, one with the AlkB demethylase [demethylase(+)] and the other without treatment [demethylase(-)]. Finally, the samples obtained were constructed using RNA Library Establishment Kit (NEB) and sequenced on an illumina NovaSeq instrument with 150bp paired end reads.