IPC분류정보
국가/구분 |
United States(US) Patent
등록
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국제특허분류(IPC7판) |
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출원번호 |
US-0633034
(2012-10-01)
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등록번호 |
US-8785127
(2014-07-22)
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발명자
/ 주소 |
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출원인 / 주소 |
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대리인 / 주소 |
Kilpatrick Townsend & Stockton LLP
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인용정보 |
피인용 횟수 :
3 인용 특허 :
79 |
초록
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The invention relates to methods and devices for analyzing single molecules, i.e., nucleic acids. Such single molecules may be derived from natural samples, such as cells, tissues, soil, air, and water without separating or enriching individual components. In certain aspects of the invention, the me
The invention relates to methods and devices for analyzing single molecules, i.e., nucleic acids. Such single molecules may be derived from natural samples, such as cells, tissues, soil, air, and water without separating or enriching individual components. In certain aspects of the invention, the methods and devices are useful in performing nucleic acid sequence analysis by probe hybridization.
대표청구항
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1. A method for determining sequence information for a polynucleotide, comprising: (a) providing an array of DNA molecules, wherein each DNA molecule comprises a fragment of said polynucleotide;(b) contacting the array with a set of informative oligonucleotide probes, wherein each probe comprises a
1. A method for determining sequence information for a polynucleotide, comprising: (a) providing an array of DNA molecules, wherein each DNA molecule comprises a fragment of said polynucleotide;(b) contacting the array with a set of informative oligonucleotide probes, wherein each probe comprises a label and a nucleotide sequence comprising the formula NxByNz, wherein: (i) each N is independently a degenerate base;(ii) each B is independently an informative base;(iii) x and z are each at least one; and(iv) y is 2 to 20;(c) detecting labeled probes once the probes have hybridized to DNA molecules in the array, thereby obtaining data indicating which of the labeled probes are hybridized to each DNA molecule; and(d) processing the data to obtain said sequence information for the polynucleotide. 2. The method of claim 1, wherein each DNA molecule comprises multiple copies of the respective fragment. 3. The method of claim 1, wherein y is 2 to 8. 4. The method of claim 1, wherein the labels are fluorescent labels. 5. The method of claim 1, wherein the DNA molecules on the array comprise overlapping fragments of an entire human genome. 6. A method for determining sequence information for a polynucleotide, comprising: (a) providing an array of DNA molecules, wherein each DNA molecule comprises a fragment of said polynucleotide;(b) contacting the array with a first set of informative oligonucleotide probes, wherein each probe comprises a label and a nucleotide sequence comprising the formula NxBy or the formula ByNz, wherein: (i) each N is independently a degenerate base;(ii) each B is independently an informative base;(iii) x and z are each at least one; and(iv) y is 2 to 20;(c) detecting labeled probes from said first set once the probes have hybridized to DNA molecules in the array, thereby obtaining data indicating which of the labeled probes are hybridized to each DNA molecule; and(d) processing the data to obtain said sequence information for the polynucleotide. 7. The method of claim 6, wherein each probe of the set comprises the formula NxBy. 8. The method of claim 6, wherein each probe of the set comprises the formula ByNz. 9. The method of claim 6, further comprising: (e) contacting the array with a second set of oligonucleotides; and(f) ligating oligonucleotide probes from the first set to oligonucleotides from the second set that are hybridized to adjacent sites on the DNA molecules on the array. 10. The method of claim 9, wherein the first set of probes is a universal probe set. 11. The method of claim 6, wherein x is 1 to 4. 12. The method of claim 6, wherein y is 2 to 8. 13. The method of claim 6, wherein y is 4 to 5. 14. The method of claim 6, wherein z is 1 to 4. 15. The method of claim 6, wherein the labels are fluorescent labels. 16. The method of claim 6, wherein each DNA molecule comprises multiple copies of the respective fragment. 17. The method of claim 6, wherein the DNA molecules on the array comprise overlapping fragments of an entire human genome. 18. A method for determining sequence information for a polynucleotide, comprising: (a) providing an array of DNA molecules, wherein each DNA molecule comprises a fragment of said polynucleotide;(b) contacting the array with a first set of informative oligonucleotide probes, wherein each probe comprises a label and a nucleotide sequence comprising the formula NxBy or the formula ByNz, wherein: (i) each N is independently a degenerate base;(ii) each B is independently an informative base; and(iii) x, y, and z are each at least one;(c) contacting the array with a second set of oligonucleotide probes, each configured to bind to a site in a DNA molecule of the array adjacent to a probe from the first set;(d) detecting labeled probes from said first set once oligonucleotide probes from both the first and the second set have hybridized to DNA molecules in the array, thereby obtaining data indicating which of the labeled probes are hybridized to each DNA molecule; and(e) processing the data to obtain said sequence information for the polynucleotide. 19. The method of claim 18, further comprising ligating an oligonucleotide probe from the first set to an oligonucleotide from the second set when hybridized to adjacent sites on a DNA molecule on the array. 20. The method of claim 18, wherein probes in the first set comprise the formula NxByNz. 21. The method of claim 18, wherein y is 2 to 8. 22. The method of claim 18, wherein the DNA molecules on the array comprise overlapping fragments of an entire human genome.
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