Patent ID: 7132235
Filing Date: 2006-11-07
Classification: C12Q

Abstract:
1. A reagent kit for detecting the presence or absence of one or more specific nucleotides at a predetermined target position in a target nucleic-acid polymer, comprising: (a) a detection primer comprising a detection-primer nucleotide sequence having a primer-extension-initiation 3′-end nucleotide which constitutes a 3′ terminal end of the detection primer, the detection-primer nucleotide sequence being complementary to a primer-hybridizing nucleotide sequence of the target nucleic-acid polymer with a nucleotide in the target nucleic-acid polymer complementary to the primer-extension-initiation 3′-end nucleotide of the detection-primer nucleotide sequence defining a primer-end complement nucleotide, the primer-hybridizing nucleotide sequence of the target nucleic-acid polymer extending towards the 3′ end of the target polymer from the primer-end complement nucleotide, the primer-end complement nucleotide being located in the target polymer at a position 3′-ward of the predetermined target position, the position of the primer-end complement nucleotide being subject to a constraint that no nucleotide of the same type as the one or more specific nucleotides to be detected be located in the target polymer in any position between the position of the primer-end complement nucleotide and the predetermined target position; (b) an enzymatic polymerizing agent; and (c) a plurality of nucleoside triphosphates including at least one deoxynucleotide and at least two different chain-terminating nucleotide analogues, at least one deoxynucleotide comprising a detectable label or an attachment moiety capable of binding a detectable label, each deoxynucleotide of said plurality of nucleoside triphosphates being complementary to a nucleotide which differs from any nucleotide to which a chain-terminating nucleotide analogue of said plurality of nucleoside triphosphates is complementary; so that in use the detection primer can hybridize to the target nucleic-acid polymer at the primer-hybridizing nucleotide sequence and form a detection-primer extension product by an enzyme-catalyzed primer-extension reaction to permit the presence or absence of a specific nucleotide at the predetermined target position to be detected by detecting the presence or absence of a corresponding detectable label in association with the detection-primer extension product.