Patent ID: 8232078
Filing Date: 2012-07-31
Classification: C07K,C12N,C12P,C12Q,C12Y

Abstract:
1. 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 DNA polymerase, wherein the DNA polymerase is joined to a sequence non-specific double-stranded nucleic acid binding domain that comprises at least 75% amino acid sequence identity to the Sso7d amino acid sequence set forth in SEQ ID NO:2 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, (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).