NUCLEIC ACID QUANTITATION METHODS
First Claim
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1. A method of determining the amount of a target nucleic acid sequence in a sample, the method comprising:
- (a) obtaining multiple distinguishable amplicons of the target nucleic acid sequence, each comprising a distinguishing tag and a target portion, wherein each target portion is complementary to an identical target nucleic acid subsequence or its complement;
(b) amplifying the amplicons in a single reaction volume; and
(c) detecting nucleic acids amplified from at least two distinguishable amplicons.
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Abstract
The invention relates to a method of determining the amount of a target nucleic acid sequence in a sample, the method comprising: obtaining multiple distinguishable amplicons of the target nucleic acid sequence, each comprising a distinguishing tag and a target portion; amplifying the amplicons in a single reaction volume; and detecting nucleic acids amplified from the amplicons. Detection of the distinguishable amplicons can be varied in each of the steps of the method, which expands the dynamic range of the nucleic acid quantification methods and improves the reliability and accuracy of the methods.
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45 Claims
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1. A method of determining the amount of a target nucleic acid sequence in a sample, the method comprising:
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(a) obtaining multiple distinguishable amplicons of the target nucleic acid sequence, each comprising a distinguishing tag and a target portion, wherein each target portion is complementary to an identical target nucleic acid subsequence or its complement; (b) amplifying the amplicons in a single reaction volume; and (c) detecting nucleic acids amplified from at least two distinguishable amplicons. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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2. A method of determining the amount of a target nucleic acid sequence in a sample, the method comprising:
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(a) obtaining multiple distinguishable amplicons of the target nucleic acid sequence, each comprising a distinguishing tag and a target portion, wherein the target portion is identical in sequence and length in each distinguishable amplicon; (b) amplifying the amplicons in a single reaction volume; and (c) detecting nucleic acids amplified from at least two distinguishable amplicons.
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35. A method of determining the amount of a target nucleic acid sequence in a sample, the method comprising:
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(a) obtaining multiple distinguishable sequencons of the target nucleic acid sequence in a single reaction volume, each comprising a distinguishing tag and a target portion, wherein each target portion is complementary to an identical target nucleic acid subsequence or its complement; and (b) detecting different amounts of least two of the sequencons, thereby increasing the dynamic range of the method of determining the amount of the target nucleic acid.
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36. A method of determining the amount of a target nucleic acid sequence in a sample, the method comprising:
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(a) obtaining multiple distinguishable amplicons of the target nucleic acid sequence, each comprising a zip code and a target portion, wherein the target portions are complementary (vs. hybridize) to an identical or overlapping target nucleic acid subsequence; (b) amplifying the amplicons in a single reaction volume; and (c) detecting nucleic acids amplified from each of the amplicons.
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37. A kit for detecting a target nucleic acid sequence in a sample comprising multiple distinguishing tags, wherein at least two of the distinguishing tags comprise:
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(a) a target binding sequence that is complementary to an identical or overlapping portion of the target nucleic acid sequence; and (b) a zip code that uniquely identifies the distinguishing tag. - View Dependent Claims (38, 39, 40, 41, 42, 43)
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44. A method for determining the amount of a target nucleic acid sequence in a sample, the method comprising:
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(a) producing multiple distinguishable amplicons in a primer-dependent enzymatic reaction, wherein at least two amplicons comprise a distinguishing tag and a target portion that is complementary to an identical or overlapping target nucleic acid subsequence or its complement; (b) differentially amplifying the amplicons in a single reaction volume; and (c) detecting nucleic acids amplified from at least two distinguishable amplicons. - View Dependent Claims (45)
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Specification