Long-Range PCR Amplification of DNA by DNA Polymerase III Holoenzyme from Thermus thermophilus

Figure 5

Assay to optimize thermostability of the Tth pol III holoenzyme. Assays were as described under Section 2. Positive controls are reactions carried out using initial buffer at 55°C. (a) Effect of different buffers in reactions on activity at 85°C. HEPES, Tris-HCl, and TAPS-Tris buffer concentrations were 25, 50, and 20 mM, respectively. (b) Assays containing different percent of glycerol tested at 87°C. (c) Assays containing the indicated concentrations of ATP were tested at 88°C. (d) Assays containing the indicated concentrations of magnesium acetate (Mg⁺⁺) were tested at 88°C. Negative control is minus Mg⁺⁺.

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