Use of continuous/contiguous stacking hybridization as a diagnostic tool
First Claim
1. A method for detecting disease associated alleles in patient genetic material comprising:
- a) immobilizing oligonucleotide molecules from a first group, at a predetermined position on a substrate, each of the molecules having a first end, a second end, and a known length, whereby the first end is attached to the substrate said oligonucleotide molecules synthesized to complement base sequences of the disease associated alleles;
b) contacting the patient genetic material with said first group of oligonucleotides to form duplexes;
c) positioning oligonucleotide molecules from a second group of oligonucleotide molecules at the second end of the immobilized oligonucleotide molecules of the first group to noncovalently extend the length of the oligonucleotide molecules of the first group, where each of the oligonucleotide molecules of the second group are tagged with a fluorochrome which radiates light at a predetermined wavelength, and where each of the oligonucleotide molecules of the second group consist of "n" number of bases and wherein just one of the bases of the oligonucleotide molecules of the second group is guanine or cytosine or thymine or adenine or uracil with the remaining bases each comprised of a universal base selected from the group consisting of 5-nitroindole, 3-nitropyrrole, and inosine; and
d) comparing the light patterns radiating from the predetermined positions on the substrate with predetermined light patterns emitted from identical substrates which were contacted with the disease associated alleles.
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Accused Products
Abstract
A method for detecting disease-associated alleles in patient genetic material is provided whereby a first group of oligonucleotide molecules, synthesized to compliment base sequences of the disease associated alleles is immobilized on a predetermined position on a substrate, and then contacted with patient genetic material to form duplexes. The duplexes are then contacted with a second group of oligonucleotide molecules which are synthesized to extend the predetermined length of the oligonucleotide molecules of the first group, and where each of the oligonucleotide molecules of the second group are tagged and either incorporate universal bases or a mixture of guanine, cytosine, thymine, and adenine, or complementary nucleotide strands that are tagged with a different fluorochrome which radiates light at a predetermined wavelength. The treated substrate is then washed and the light patterns radiating therefrom are compared with predetermined light patterns of various diseases that were prepared on identical substrates.
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Citations
20 Claims
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1. A method for detecting disease associated alleles in patient genetic material comprising:
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a) immobilizing oligonucleotide molecules from a first group, at a predetermined position on a substrate, each of the molecules having a first end, a second end, and a known length, whereby the first end is attached to the substrate said oligonucleotide molecules synthesized to complement base sequences of the disease associated alleles; b) contacting the patient genetic material with said first group of oligonucleotides to form duplexes; c) positioning oligonucleotide molecules from a second group of oligonucleotide molecules at the second end of the immobilized oligonucleotide molecules of the first group to noncovalently extend the length of the oligonucleotide molecules of the first group, where each of the oligonucleotide molecules of the second group are tagged with a fluorochrome which radiates light at a predetermined wavelength, and where each of the oligonucleotide molecules of the second group consist of "n" number of bases and wherein just one of the bases of the oligonucleotide molecules of the second group is guanine or cytosine or thymine or adenine or uracil with the remaining bases each comprised of a universal base selected from the group consisting of 5-nitroindole, 3-nitropyrrole, and inosine; and d) comparing the light patterns radiating from the predetermined positions on the substrate with predetermined light patterns emitted from identical substrates which were contacted with the disease associated alleles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for determining nucleotide sequences from patient genetic material comprising:
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a) immobilizing oligonucleotide molecules from a first group, at a predetermined position on a substrate, each of the molecules having a first end, a second end, and a known length, whereby the first end is attached to the substrate, said oligonucleotide molecules synthesized to complement base sequences of the disease associated alleles; b) contacting the patient genetic material with said first group of oligonucleotides to form duplexes; c) positioning oligonucleotide molecules from a second group of oligonucleotide molecules at the second end of the immobilized oligonucleotide molecules of the first group to noncovalently extend the length of the oligonucleotide molecules of the first group, where each of the oligonucleotide molecules of the second group are tagged with a fluorochrome which radiates light at a predetermined wavelength, and where each of the oligonucleotide molecules of the second group consist of "n" number of bases and wherein just one of the bases of the oligonucleotide molecules of the second group is guanine or cytosine or thymine or adenine and the remaining bases are each comprised of a universal base selected from the group consisting of 5-nitroindole, 3-nitropyrrole, and inosine; and d) repeating step c to determine the complete sequence of the patient genetic material duplexed with both the oligonucleotide molecules from the first group and the oligonucleotide molecules from the second group. - View Dependent Claims (11, 12, 13, 14)
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15. A method for determining nucleotide sequences from patient genetic material comprising:
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a) immobilizing oligonucleotide molecules from a first group, at a predetermined position on a substrate, each of the molecules having a first end, a second end, and a known length, whereby the first end is attached to the substrate, said nucleotide molecules synthesized to complement base sequences of the disease associated alleles; b) contacting the patient genetic material with said first group of oligonucleotides to form duplexes; c) positioning oligonucleotide molecules from a second group of oligonucleotide molecules at the second end of the immobilized oligonucleotide molecules of the first group to noncovalently extend the length of the oligonucleotide molecules of the first group, where each of the oligonucleotide molecules of the second group are tagged with a fluorochrome which radiates light at a predetermined wavelength, and where each of the oligonucleotide molecules of the second group is a pentamer containing four universal bases and just one base is guanine or cytosine or thymine or adenine; and d) repeating step c to determine the base sequence of the patient genetic material that extends five bases beyond the second end of the immobilized oligonucleotide molecule. - View Dependent Claims (16, 17, 18, 19, 20)
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Specification