Patent ID: 7670808
Filing Date: 2010-03-02
Classification: C07K,C12N,C12P,C12Y,Y02P

Abstract:
1. A method of amplifying a 5kb or longer subsequence of a target nucleic acid in an aqueous solution using a polymerase chain reaction, the method comprising: (i) contacting the target nucleic acid with a DNA polymerase comprising a sequence-non-specific double-stranded nucleic-acid-binding domain that comprises an amino acid sequence that has at least 75% identity to the amino acid sequence of SEQ ID NO:2 joined to a DNA polymerase domain with error-correcting activity, where the sequence non-specific double-stranded nucleic-acid-binding domain enhances the processivity of the DNA polymerase domain compared to an identical DNA polymerase domain not having the sequence non-specific double-stranded nucleic acid binding domain; and wherein the solution 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 DNA polymerase domain to extend a primer that is hybridized to the target nucleic acid sequence to a length of 5 kb or longer; (ii) incubating the solution using a polymerase chain reaction temperature profile that amplifies the 5 kb or longer subsequence.