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Example of a steady-state kinetics experiment to determine the kinetics of single nucleotide insertion opposite the 10-mer peptide cross-linked to C7 of 7-deaza-G. Single nucleotide incorporation assays were performed using hPol η and 10-mer peptide-deaza-G cross-links in the presence of increasing concentrations of individual dTTPs. Polymerization reactions were quenched at preselected time points and loaded onto 20% (w/v) denaturing polyacrylamide gels.
