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Effects of PCNA mutants on pol δ-catalyzed elongation of poly dA:oligo dT in the absence of RFC. Reaction mixtures (15 μl) contained 40 mM bicine-Tris buffer (pH 6.8), 60 μg/ml BSA, 1 mM DTT, 6 mM magnesium acetate, 30 μM [α32P]dTTP (5,200 cpm/pmol), 12.5 μM poly(dA)4000:oligo(dT)12–18 (1:20), PCNA (in amounts as indicated), and 0.15 units of hpol δ. Reactions were incubated for 20 min at 37°C, and aliquots were used to determine the amount of poly dT synthesized.
