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In vitro pseudouridylation of U2 snRNA transcript by using recombinant Pus1p pseudouridine synthase. After incubation of the transcript with the purified Pus1p enzyme (as described in Materials and Methods), the phenol-extracted RNA was analyzed by the CMCT-RT method (lanes 1, 2, 3, and 4) as described in the legend to Fig. 1. The strong alkaline-resistant RT stop at position 44 is indicated by an arrow. Lanes U, G, C, and A correspond to the sequencing ladder.
