Analying polynucleotide sequences
DCFirst Claim
1. A method of making an array of oligonucleotides, which comprises:
- attaching a plurality of oligonucleotides to an impermeable surface of a support, the oligonucleotides having different predetermined sequences and being attached at different known locations on the surface of the support through a computer-controlled printing device.
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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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Citations
12 Claims
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1. A method of making an array of oligonucleotides, which comprises:
attaching a plurality of oligonucleotides to an impermeable surface of a support, the oligonucleotides having different predetermined sequences and being attached at different known locations on the surface of the support through a computer-controlled printing device. - View Dependent Claims (2)
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3. A method for constructing an array of oligonucleotides of length s and composed of different nucleotides, which method comprises:
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a) applying precursors for the different nucleotides separately to a plurality of different regions of a surface, b) applying precursors for the different nucleotides separately to a plurality of different regions amongst the plurality of different regions defined in a), c) repeating the process until each of said regions contains oligonucleotides of length s. - View Dependent Claims (4, 5, 6, 7, 8)
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9. A method of analysing a polynucleotide, which method comprises:
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applying a labelled polynucleotide to be analysed or fragments thereof to an array of oligonucleotides under hybridisation conditions, wherein the array comprises a support having an impermeable surface to which a plurality of oligonucleotides having different predetermined sequences are attached to different known regions on the surface, and analysing the polynucleotide by observing the regions where the polynucleotide or fragment thereof hybridizes and the regions where the polynucleotide or fragment thereof does not hybridize.
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10. A method of comparing polynucleotide sequences, which method comprises:
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applying the polynucleotides to an array of oligonucleotides under hybridizing conditions, wherein the oligonucleotides have different predetermined sequences and are attached at different known locations on an impermeable surface of a support, and observing the differences between the patterns of hybridisation.
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11. A method for analysing multiple sequences in multiple polynucleotides, which comprises:
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a) laying down stripes of oligonucleotides corresponding to each sequence on the surface of an impermeable solid support, b) applying the polynucleotides to the surface under hybridisation conditions in stripes orthogonal to those of the oligonucleotides, c) observing hybridisation at a site of intersection as an indication of the presence of a sequence in the polynucleotide.
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12. An array of oligonucleotides comprising a support having an impermeable surface to which a plurality of oligonucleotides are attached, the oligonucleotides having different nucleotide sequences and being attached at different known locations on the surface of the support, wherein each oligonucleotide has the sequence Nm AATAAANn or its complement where N is any nucleotide residue and m+n is at least about 8.
Specification