Figure 2.

Extension of RNA primer by P. furiosus family B DNA polymerase. The extension of RNA primers annealed to complementary DNA templates was determined in a buffer consisting of 20 mM Tris–HCl, pH 8.8, 10 mM (NH₄)₂SO₄, 10 mM KCl, 2 mM MgSO₄, 0.1% Triton X-100, 100 ng/μl BSA, 100 μM dNTPs and 4 U Rnsin. Approximately 50 nM of RNA primer–DNA template substrates was incubated with 100 nM Pfu DNA polymerase at 50°C for 0, 1, 2, 4, 8, 15, 30 and 60 min (A) or 30 min (B and C). Different RNA primers and DNA templates were annealed to form the matched and mismatched RNA/DNA hybrids used in the extension reactions. Lowercase and uppercase denote RNA and DNA, respectively.