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US 5,902,723 AGranted
Analysis of surface immobilized polymers utilizing microfluorescence detection
OwnerWilliam J. Dower, Stephen P. A. Fodorinventors
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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 monomers on solid phase immobilized polymers. Reagents which specifically recognize terminal manomers are used to label polymers at defined positions on a solid substrate.
Claims15
A method of analyzing a plurality of nucleic acids, comprising:
- providing a plurality of different nucleic acid sequences immobilized upon a solid support, each of said different sequences being attached to said solid support in a distinct position:
- treating said plurality of nucleic acid sequences with a catalytic antibody that is capable of catalyzing removal of a specific nucleotide in a nucleotide-specific manner to selectively remove a single terminal nucleotide;
- identifying positions on said solid support in which a terminal nucleotide has been removed;
- and
- repeating said treating and identifying steps to analyze said sequences of said plurality of different nucleic acids.
2 more elements
A method of polynucleotide analysis, comprising:
- (a) contacting at least one polynucleotide with a surface of a solid substrate, the surface having attached thereto in spatially defined locations a plurality of different oligonucleotide primers of defined nucleotide sequence and length, wherein the oligonucleotides are patterned on the surface of the substrate such that the oligonucleotide primers with different nucleotide sequences are located in different locations;
- (b) hybridizing the oligonucleotide primers to at least one region of complementary nucleotide sequence contained in the at least one polynucleotide in a reaction mixture comprising a nucleic acid polymerase and a labelled nucleotide under conditions whereby the at least one hybridized primer is elongated by template-directed addition of the labelled nucleotide;
- and
- (c) determining the location of the at least one hybridized primer from the location of label, the location of the hybridized primer indicating its sequence and thereby the complementary sequence of the at least one polynucleotide.
1 more element
A method for determining the incorporation of nucleotides or polynucleotides comprising:
- providing a nucleic acid array, said array having associated therewith at least two different nucleic acids, each of said different nucleic acids being associated with said array in a different defined position;
- hybridizing at least a portion of a target nucleic acid to said nucleic acid array;
- contacting said nucleic acid array with a plurality of nucleotides or polynucleotides and a template dependent nucleic acid polymerase;
- determining the incorporation of said nucleotides or polynucleotides at said different defined positions.
1 more element
Dependencies are inferred from the claim text.