To perform an 8-OHdG quantitation assay (with DNA extracted from eukaryotic cells as samples), an enzyme that digests DNA down to the single nucleotide level is needed. Nuclesase P1 is recommended by the manufacturer, but is there any alternative that can guarantee the same results?This is a critical step in this assay, because even the presence of dinucleotides in the sample will affect the results.
Pay attention to S1 (acid pH 4.8) or mung bean nuclease (pH 7)!
Additionally, you can use combined treatment with DNAse I and snake venom PDE I.
In all cases you need to do control experiments to find optimal conditions. HPLC can be in help.
Good luck!
Yuri F. Drygin,
In my understanding, nuclease S1 only digests partially DNA. Maybe in combination with other enzymes could work.
Thank you for your response!
S1 nuclease hydrolyses DNA completely. Hydrolysis by DNAse I dropps down to zero for (4-6) oligomers.
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