Methods of using improved polymerases

A method of performing a quantitative real-time polymerase chain reaction on a target nucleic acid present in a solution that comprises a DNA-binding fluorescent dye, the method comprising:

• (a) contacting the target nucleic acid with a thermally stable DNA polymerase, wherein the DNA polymerase is joined to a sequence non-specific double-stranded nucleic acid binding domain that specifically binds to polyclonal antibodies generated against either Sso7d (SEQ ID NO:2) or Sac7d (amino acids 7-71 of SEQ ID NO:10) and enhances the processivity of the DNA polymerase compared to an identical DNA polymerase not having the sequence non-specific double-stranded nucleic-acid binding domain fused to it,
• wherein the solution comprises the DNA-binding fluorescent dye, which exhibits altered fluorescence emissions when the dye is bound to double-stranded DNA, and is of a composition that permits the sequence non-specific double-stranded nucleic acid binding domain to bind to the target nucleic acid and the polymerase domain to extend a primer that is hybridized to the target nucleic acid sequence;
• (b) incubating the solution under conditions in which the primer is extended by the DNA polymerase,
• (c) exposing the solution to a suitable excitation light and measuring fluorescence emission from the DNA-binding fluorescent dye and;
• (d) performing at least one additional cycle of amplification comprising steps (a) to (c).