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Assay Method Information |
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| | DNA Polymerase Assay |
| Description: | For the DNA polymerase reaction, the dehydroaltenusin derivative was dissolved in dimethyl sulfoxide (DMSO) at various concentrations and sonicated for 30 s. Four microlitres of each sonicated sample were mixed with 16 uL of enzyme (final 0.05 units) in reaction buffer, and kept at 0 deg C for 10 min. These inhibitor-enzyme mixtures were added to 16 uL of each enzyme standard reaction mixture containing DNA template-primer and nucleotide, and incubated at 37 deg C for 60 min. Activity without the inhibitor was considered 100%, and the remaining activity at each concentration of the inhibitor was determined relative to this value. One unit of pol activity was defined as the amount of enzyme that catalyzed the incorporation of 1 nmol of dNTP (i.e., dTTP) into synthetic DNA template-primer in 60 min at 37 deg C under the normal reaction conditions. |
| Affinity data for this assay | |
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