The formation and repair of DNA damage, and its toxicological influences, can strongly depend on the DNA sequence in which they occur. Sequence-specific methods for DNA damage quantification could provide missing information regarding DNA repair and mutagenesis mechanisms, and more precise data about the effective dose of DNA damaging agents. We developed a fast, simple and robust fluorescence-based assay for evaluating the kinetics of dNTP incorporation by a lesion-bypass DNA polymerase in a single nucleotide primer-extension reaction. This allowed us to determine the incorporation efficiency for DNA polymerase IV (Dpo4) of a small panel of synthetic dNTPs opposite O6-methyl-guanine-containing templates and to assess their potential as probes for evaluating O6-methyl-guanine.

Incorporation efficiency of nucleoside probes opposite the mutagenic DNA adduct <i>O</i><sup>6</sup>-methyl-guanosine evaluated with a new enzymatic fluorescence read-out assay

244th National Meeting (2012)

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Incorporation efficiency of nucleoside probes opposite the mutagenic DNA adduct O6-methyl-guanosine evaluated with a new enzymatic fluorescence read-out assay

Incorporation efficiency of nucleoside probes opposite the mutagenic DNA adduct O⁶-methyl-guanosine evaluated with a new enzymatic fluorescence read-out assay