AzaIP-seq

All characterized mammalian m5C-RNA methyltransferases form a reversible covalent intermediate with their cytosine substrate that is trapped when conducted on the cytosine analog 5-azacytidine. We pretreated human U87MG cells which was transfected with NSUN5 with 5-azacytidine or DMSO, and then precipitated its RNA with anti-Flag beads as our sequencing sample. The aim of this experiment is to figure out which RNA that NSUN5 binds to.