Apparatus and method for analyzing polynucleotide sequences and method of generating oligonucleotide arrays
First Claim
1. A method for generating an array of oligonucleotides of chosen lengths within discrete cells of a support material comprising the steps ofa) segregating a support material into discrete cell locations;
- b) coupling a nucleotide precursor to a first set of cell locations;
c) coupling a nucleotide precursor to a second set of cell locations;
d) coupling a nucleotide precursor to a third set of cell locations;
e) and continuing the sequence of coupling steps until the desired array has been generated,the coupling being effected at each location either to the surface of the support or to a nucleotide coupled in a previous step at the location.
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Abstract
This invention provides an apparatus and method for analyzing a polynucleotide sequence; either an unknown sequence or a known sequence. A support, e.g. a glass plate, carries an array of the whole or a chosen part of a complete set of oligonucleotides which are capable of taking part in hybridization reactions. The array may comprise one or more pair of oligonucleotides of chosen lengths. The polynucleotide sequence, or fragments thereof, are labelled and applied to the array under hybridizing conditions. Applications include analyses of known point mutations, genomic fingerprinting, linkage analysis, characterization of mRNAs, mRNA populations, and sequence determination.
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8 Claims
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1. A method for generating an array of oligonucleotides of chosen lengths within discrete cells of a support material comprising the steps of
a) segregating a support material into discrete cell locations; -
b) coupling a nucleotide precursor to a first set of cell locations; c) coupling a nucleotide precursor to a second set of cell locations; d) coupling a nucleotide precursor to a third set of cell locations; e) and continuing the sequence of coupling steps until the desired array has been generated, the coupling being effected at each location either to the surface of the support or to a nucleotide coupled in a previous step at the location. - View Dependent Claims (2, 3, 4, 5)
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6. A method of analyzing a polynucleotide, by the use of apparatus comprising a support segregated into at least two defined cells, each cell having attached thereto oligonucleotides containing predetermined sequences, the oligonucleotides of each cell having been synthesized in situ and being attached to the surface of the support through a covalent linkage, where the sequence of the oligonucleotides of a first cell is different than the sequence of the oligonucleotides of a second cell,
which method comprises applying the polynucleotide under hybridization conditions to the support and observing the location of hybridized polynucleotide on the support, wherein the oligonucleotides are attached to the surface as an array of parallel stripes, and at least two polynucleotides are analyzed simultaneously by applying the polynucleotides to the array in the form of separate stripes orthogonal to the oligonucleotides stripes.
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7. A method of analyzing a polynucleotide, by the use of apparatus comprising a support segregated into at least two defined cells, each cell having attached thereto oligonucleotides containing predetermined sequences, the oligonucleotides of each cell having been synthesized in situ and being attached to the surface of the support through a covalent linkage, where the sequence of the oligonucleotides of a first cell is different than the sequence of the oligonucleotides of a second cell,
which method comprises randomly degrading the polynucleotide to form a mixture of oligomers, the mixture being labeled to form labeled material, applying the labeled material under hybridization conditions to the support and observing the location of the labeled material on the support.
Specification