Sequencing of surface immobilized polymers utilizing microflourescence detection
First Claim
Patent Images
1. A method for determining the incorporation of nucleotides comprising:
- (a) providing a plurality of target template nucleic acids attached to a substrate in positionally separated regions at a density of at least 1000 regions per cm2;
(b) hybridizing at least one primer to said plurality of target template nucleic acids to form hybridized primer;
(c) contacting said hybridized primer with, at least one nucleotide and a template-dependent nucleic acid polymerase, wherein said nucleotide comprises a label, under conditions whereby the hybridized primer is elongated by template-directed addition of a nucleotide; and
(c) detecting the addition of said nucleotide by detecting the label and thereby determining the incorporation of the nucleotide.
0 Assignments
0 Petitions
Accused Products
Abstract
Means for simultaneous parallel sequence analysis of a large number of biological polymer macromolecules. Apparatus and methods may use fluorescent labels in repetitive chemistry to determine terminal monomers on solid phase immobilized polymers. Reagents which specifically recognize terminal monomers are used to label polymers at defined positions on a solid substrate.
357 Citations
13 Claims
-
1. A method for determining the incorporation of nucleotides comprising:
-
(a) providing a plurality of target template nucleic acids attached to a substrate in positionally separated regions at a density of at least 1000 regions per cm2; (b) hybridizing at least one primer to said plurality of target template nucleic acids to form hybridized primer; (c) contacting said hybridized primer with, at least one nucleotide and a template-dependent nucleic acid polymerase, wherein said nucleotide comprises a label, under conditions whereby the hybridized primer is elongated by template-directed addition of a nucleotide; and (c) detecting the addition of said nucleotide by detecting the label and thereby determining the incorporation of the nucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 8)
-
-
7. A method for polynucleotide analysis comprising:
-
(a) providing a plurality of different target template nucleic acids attached to a support in positionally separated regions at a density of at least 1000 regions per cm2; (b) hybridizing a primer to the target template nucleic acids to obtain a hybridized primer; (c) contacting the hybridized primer with a plurality of species of nucleotides and a template dependentnucleic acid polymerase, wherein the nucleotides comprise a removable label and a removable blocking group and wherein each species of nucleotides comprises a different label that is distinguishable from the label on each of the other species of nucleotides; (d) extending the hybridized primer by incorporation of a nucleotide; (e) scanning the substrate to detect a signal from the label, wherein the signal indicates the species of nucleotide that was incorporated in (d); (f) removing the label and the blocking group; (g) repeating steps (c) to (f) at least once; and (h) determining at least a portion of the sequence of the target template nucleic acid from the species of nucleotide incorporated in step (d). - View Dependent Claims (9, 10, 11, 12, 13)
-
Specification