Patent ID: 7700281

Claim:
A method of duplicating a template nucleic acid, or a portion thereof, wherein a primer having a nucleotide sequence that is complementary to a target portion of the template nucleic acid is hybridized to the template nucleic acid and then extended via an enzyme, the method comprising the steps of: (a) at a first temperature, preparing a reaction mixture comprising a primer, a template nucleic acid, an enzyme effective to catalyze primer extension and single-stranded nucleic acid binding protein comprising at least one of wild-type T7 gp2.5 and its mutant variants, or a combination thereof, wherein said single-stranded nucleic acid binding protein inhibits the generation of nonspecific primer extension products at said first temperature, (b) at a second temperature higher than said first temperature, carrying out a hybridization reaction in said reaction mixture to produce a hybridized product, wherein at said second temperature said primer is substantially uninhibited by said single-stranded nucleic acid binding protein from participating in the hybridization reaction, and (c) at a third temperature higher than said first temperature, carrying out a primer extension reaction to produce from said hybridized product an extended product; said primer becoming uninhibited from participating in at least one of the hybridization and extension reactions as a result of or in conjunction with said single-stranded nucleic acid binding protein being denatured at a temperature above said first temperature.