Patent ID: 7871771
Filing Date: 2011-01-18
Classification: C12Q

Abstract:
1. A method of characterizing a target region of a nucleic acid template, comprising: a) initiating a nucleic acid polymerization reaction on a support, by forming a reaction mixture, said reaction mixture including said nucleic acid template, a primer, a nucleic acid polymerizing enzyme, and at least one terminal-phosphate-labeled nucleotides each containing a distinct label, wherein a component of said reaction mixture or a first complex of two or more of said components, is immobilized on said support, and said component or components are selected from the group consisting of said nucleic acid template, said primer, and said nucleic acid polymerizing enzyme, and wherein each of said at least one terminal-phosphate-labeled nucleotides contains a base complementary to each of the four naturally occurring bases; b) progressing said nucleic acid polymerization reaction by incubating said reaction mixture, in the absence of a divalent cation, to form a second complex comprising said nucleic acid template, said primer, said nucleic acid polymerizing enzyme, and a terminal-phosphate-labeled nucleotide, wherein said terminal-phosphate-labeled nucleotide contains a base complementary to the template base at the site of polymerization; c) removing unbound terminal-phosphate-labeled nucleotides and other components of said reaction mixture; and d) detecting the label of said terminal-phosphate-labeled nucleotide from said second complex to characterize said target region of said nucleic acid template.