Bisulfite Incorporation Hindered Ligation-based Method for Locus-Specific Detection and Quantification of Pseudouridine

We applied BIHIND-seq to 15 sites of Hela 18S rRNA to probe Pseudouridine modification at the target sites and checked how pseudouridine level changed after DKC1 knockdown. DNA probes were annealed to Hela 18S rRNA with or without DKC1 KD. The annealed product was treated with Bst 2.0 and SplintR ligase. The final ligated product was subjected to PCR amplification for library preparation.