Patent ID: 7846695
Filing Date: 2010-12-07
Classification: C12N,C12Q

Abstract:
1. A method for amplifying a polynucleotide comprising the steps of mixing the following elements a) through f), and incubating under conditions that enable a complementary strand synthesis reaction accompanying strand displacement: a) a first primer that (i) provides at its 3′-end a starting point for complementary strand synthesis to a region that defines the 3′-side of one of the strands that compose a target nucleotide sequence, and (ii) has on its 5′-side a nucleotide sequence that is complementary to the arbitrary region of a complementary strand synthesis reaction product that uses this primer as a starting point; b) a second primer that has (i) on its 3′-end a nucleotide sequence that provides a starting point for complementary strand synthesis to a region that defines the 3′-side of a target nucleotide sequence in an elongation product that uses the first primer as a starting point, and (ii) on its 5′-side a nucleotide sequence that is complementary to the arbitrary region of a complementary strand synthesis reaction product that uses this primer as a starting point; c) a first loop primer that provides a starting point for complementary strand synthesis using an elongation product of the first primer as a template, wherein the starting point of the first loop primer is different from the starting points of the first and second primers, and the starting point of the first loop primer is between a region corresponding to the first primer sequence in the elongation product of the first primer and the arbitrary region which anneals to the first primer; d) a DNA polymerase catalyzing complementary strand synthesis accompanying strand displacement; e) a substrate for complementary strand synthesis; and, f) a test polynucleotide comprising a target nucleotide sequence.