Analysis of surface immobilized polymers utilizing microfluorescence detection
DCFirst Claim
1. A method for determining the positions of polynucleotides which terminate with a given monomer, comprisingproviding a plurality of different polynucleotides attached at distinct positions of a surface of a support by annealing of the polynucleotides to oligonucleotides immobilized to regions of the surface of the support at a density of at least 1000 regions/cm2, the polynucleotides having terminal monomers labeled in a monomer-type specific manner;
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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 manomers on solid phase immobilized polymers. Reagents which specifically recognize terminal manomers are used to label polymers at defined positions on a solid substrate.
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Citations
15 Claims
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1. A method for determining the positions of polynucleotides which terminate with a given monomer, comprising
providing a plurality of different polynucleotides attached at distinct positions of a surface of a support by annealing of the polynucleotides to oligonucleotides immobilized to regions of the surface of the support at a density of at least 1000 regions/cm2, the polynucleotides having terminal monomers labeled in a monomer-type specific manner; - and
scanning said surface, thereby determining the positions of said labeled monomer. - View Dependent Claims (9, 10, 14)
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2. A method for concurrently determining which subset of a plurality of different polymers attached at distinct positions of a surface of a solid substrate at separable locations at a density of at least 1000 locations per cm2 terminates with a given terminal subunit, said method comprising the steps of:
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mixing said solid substrate with a solution comprising a reagent, and thereby selectively marking positionally distinct polymers which terminate with said given terminal subunit; and determining with a detector which separable locations are marked, thereby determining which subset of said positionally distinct polymers terminated with said given terminal subunit. - View Dependent Claims (3, 4)
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5. A method for determining which subset of a plurality of primer polynucleotides have a predetermined oligonucleotide, wherein the polynucleotides are complementary to and annealed to different template strands which are attached at distinct positions of a surface of a solid substrate at a density of at least 1000 locations per cm2, said method comprising the steps of:
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selectively marking said subset of primer polynucleotides having the predetermined oligonucleotide; and detecting which polynucleotides are marked. - View Dependent Claims (6, 7, 8)
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11. A method for determining a subset of polynucleotides which terminate with a given monomer, comprising
providing a plurality of different polynucleotides attached at different positions of a surface of a support by annealing of the polynucleotides to oligonucleotides immobilized to regions of the surface at a density of at least 1000 regions per cm2, the polynucleotides having terminal monomers labeled: -
in a monomer type specific manner; and scanning said surface, thereby determining the subset of polynucleotides terminate with a given monomer. - View Dependent Claims (12, 13, 15)
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Specification