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Enrichment test of the 5hmU pull-down assay. The 75-nt-long DNA containing T, 5hmU, 5hmC, or 5mC was added to trypanosome DNA as spike-in controls and the JGT reaction and J-IP was performed as described in the “Materials and methods” section. The 5hmU DNA substrate analyzed without the JGT reaction (5hmU no JGT) is provided as a negative control. The %IP was calculated relative to input DNA. All experiments were performed in triplicate, error bars represent standard deviations.
