Variation analysis for multiple templates on a solid support
First Claim
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1. A method of detecting molecules, said method comprising the steps of:
- (a) providing a solid support comprising molecules associated with a site on the solid support such that the molecules are detected in aggregate during a detection step, wherein the site comprises at least two different types of molecules;
(b) detecting a signal corresponding to the aggregate of molecules at the site;
(c) providing an initial estimate of the amount of signal corresponding to different types of molecules at the site;
(d) calculating the amount of signal corresponding to different types of molecules at the site using the initial estimate, thereby obtaining a signal estimate; and
(e) iteratively updating the initial estimate until the signal estimates converge, thereby detecting molecules associated with the site.
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Abstract
Disclosed herein are compositions and methods used for detecting different types of molecules associated with a site on a solid support.
34 Citations
21 Claims
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1. A method of detecting molecules, said method comprising the steps of:
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(a) providing a solid support comprising molecules associated with a site on the solid support such that the molecules are detected in aggregate during a detection step, wherein the site comprises at least two different types of molecules; (b) detecting a signal corresponding to the aggregate of molecules at the site; (c) providing an initial estimate of the amount of signal corresponding to different types of molecules at the site; (d) calculating the amount of signal corresponding to different types of molecules at the site using the initial estimate, thereby obtaining a signal estimate; and (e) iteratively updating the initial estimate until the signal estimates converge, thereby detecting molecules associated with the site. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of identifying a target region of a nucleic acid, said method comprising:
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(a) associating a first subpopulation of nucleic acids with a site on a solid support, wherein each of the nucleic acids of said first subpopulation comprise a target region that is identical to the target region of the other nucleic acids of the first subpopulation; (b) associating a second subpopulation of nucleic acids with the site on the solid support, wherein each of the nucleic acids of said second subpopulation comprise an identical target region that is a variant of the target region of the nucleic acids of said first subpopulation; (c) detecting a signal corresponding to one or more nucleotides of the target region of first subpopulation nucleic acids and one or more nucleotides of the variant of the target region of second subpopulation nucleic acids; (d) estimating the fraction of first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site; (e) calculating the fraction of first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site using the fraction estimate; and (f) iteratively updating the fraction estimate until the estimates converge, thereby identifying a target region of a nucleic acid.
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19. A method for identifying a biosignature, said method comprising the steps of:
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(a) providing samples obtained from a plurality of subjects, wherein the samples comprise molecules; (b) tagging molecules from the samples so as to identify the subject from which each sample originated; (c) associating molecules from the samples with a site on a solid support such that the molecules are detected in aggregate during a detection step, wherein the site comprises at least two different types of molecules; (d) obtaining a biosignature for molecules associated with the site by; i) detecting a signal corresponding to the aggregate of the molecules at the site; ii) estimating the fraction of different types of molecules at the site or the amount of signal corresponding to different types of molecules at the site; iii) calculating the amount of signal corresponding to different types of molecules at the site using the fraction estimate, or calculating the fraction of different types of molecules at the site using the signal estimate; and iv) iteratively updating the fraction estimate and signal estimate until the estimates converge, thereby obtaining a biosignature for molecules at the site; and (e) comparing the biosignature obtained in step (d) to a reference biosignature, thereby identifying said bio signature.
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20. A method of identifying a target region of a nucleic acid, said method comprising:
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(a) associating a first subpopulation of nucleic acids with a site on a solid support, wherein each of the nucleic acids of said first subpopulation comprise a target region that is identical to the target region of the other nucleic acids of the first subpopulation; (b) associating a second subpopulation of nucleic acids with the site on the solid support, wherein each of the nucleic acids of said second subpopulation comprise an identical target region that is a variant of the target region of the nucleic acids of said first subpopulation; (c) detecting a signal corresponding to one or more nucleotides of the target region of first subpopulation nucleic acids and one or more nucleotides of the variant of the target region of second subpopulation nucleic acids; (d) estimating the amount of signal corresponding to first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site; (e) calculating the fraction of first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site using the signal estimate; and (f) iteratively updating the signal estimate until the estimates converge, thereby identifying a target region of a nucleic acid.
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21. A method of identifying a target region of a nucleic acid, said method comprising:
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(a) associating a first subpopulation of nucleic acids with a site on a solid support, wherein each of the nucleic acids of said first subpopulation comprise a target region that is identical to the target region of the other nucleic acids of the first subpopulation; (b) associating a second subpopulation of nucleic acids with the site on the solid support, wherein each of the nucleic acids of said second subpopulation comprise an identical target region that is a variant of the target region of the nucleic acids of said first subpopulation; (c) detecting a signal corresponding to one or more nucleotides of the target region of first subpopulation nucleic acids and one or more nucleotides of the variant of the target region of second subpopulation nucleic acids; (d) estimating the fraction of signal corresponding to first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site; (e) calculating the signal of first subpopulation nucleic acids and second subpopulation nucleic acids associated with the site using the fraction estimate; and (f) iteratively updating the fraction estimate until the estimates converge, thereby identifying a target region of a nucleic acid.
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Specification