Supplementary Figure 4: Bypass of a single rNMP by Phusion DNA polymerase
From: Ribose-seq: global mapping of ribonucleotides embedded in genomic DNA
5′-radiolabeled 30-nt primer, ByPrim (Supplementary Table 8), was annealed to the 46-nt template oligo containing either rCMP (ByTemp.rC) or rUMP (ByTemp.rU) in the 8th position. 100 nM of annealed substrate was incubated with 0.2 units of Phusion High-Fidelity DNA Polymerase (NEB) and 2 mM dNTPs in appropriate buffer (see Methods) for 30 sec at 72 °C. The reactions were quenched and analyzed by urea-PAGE. Median bypass probabilities from four independent reactions are shown. See Supplementary Table 5 for more statistics. First left lane, ss DNA ladder. The primer extension assay showed no significant difference between bypass efficiency over rUMP and rCMP by Phusion DNA polymerase (Supplementary Table 5).
