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Assay Method Information

Assay Name:  DNA Pol β PEX Assay
Description:  Pol β reactions were made in a final volume of 20uL containing 50mM Tris-HCl pH 7.9, 20mM KCl, 5% (v/v) glycerol, 1mM DTT, 2mM MnCl2, 250nM purified recombinant pol β, 150nM radioactive-labeled primer F20H, 225nM template F33A, 0.1mg/mL BSA, 1uL compound solution with various concentrations in DMSO (1uL DMSO for the solvent control). 5uL dNTP (15uM final) was added to initiate the reactions and positive controls. Reactions and controls were incubated for 30 min at 37 degrees Celsius and subsequently quenched using 45uL PAGE loading solution (80% (v/v) formamide, 20mM EDTA, 0.05% (w/v) bromophenol blue, 0.05% (w/v) xylene cyanol) per well. The reaction mixture was denatured for 5 min at 95 degrees Celsius and analyzed by 12% PAGE containing 8M urea.
Affinity data for this assay
 

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Last update November 1, 2007
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