Fig. 1. Peroxotungstate reaction on hm5C-containing RNA. (a) Illustration of the peroxotungstate reaction and workflow of WO-Seq. Hm5C-containing RNA is specifically oxidized by peroxotungstate, and then reverse transcribed by thermostable group II intron reverse transcriptase (TGIRT). The oxidation product of hm5C (thT) is converted to T during cDNA synthesis, thus can be used for base-resolution sequencing of hm5C in RNA. (b) HPLC-MS/MS results of the hydrolysed product of synthesized hm5C-containing RNA1 before and after the peroxotungstate reaction. Peaks of adenosine (rA), guanosine (rG) and hm5C are labelled in the figure. (c) MALDI-MS characterization of an hm5C-containing RNA fragment of RNA1 treated with peroxotungstate. Calculated m/z is shown in black, observed m/z is shown in red.
