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  • The quantitative HydraPsiSeq protocol for pseudouridine mapping and quantification is based on random (statistical) cleavage at all uridine residues in RNA by combination of hydrazine treatment (ring-opening of the pyrimidine base in uridines) and aniline (cleavage of the resulting RNA abasic site and release of 5′-phosphorylated RNA fragments directly competent for adapter ligation at the N+1 nucleotide in the sequence, Figure 1A, Supplementary Figure S1).
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