Determining nucleic acid fragmentation status by coincident detection of two labeled probes
First Claim
1. A method of determining fragmentation status of a nucleic acid of interest in a sample at the start of the method, the method comprising:
- contacting the nucleic acid of interest in a reaction mixture with two or more different probes complementary to sites separated by a point of potential fragmentation, wherein the probes each comprise a detectable marker;
flowing the nucleic acid of interest into a detection region;
detecting detectable marker signals from the probes; and
determining fragmentation status of the nucleic acid of interest, wherein coincident detection of signals from two or more of the different probes indicates the nucleic acid of interest was not fragmented between the probes at the start of the method and has at least a given length, and wherein detecting a signal from only one of the different probes indicates the nucleic acid of interest was fragmented.
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Abstract
Methods for determining nucleic acid fragmentation status are provided. A nucleic acid of interest in a reaction mixture is contacted with two or more different probes complementary to sites separated by a point of potential fragmentation. The probes each comprise a detectable marker. The nucleic acid of interest is flowed into a detection region, where two or more coincident detectable marker signals from the probes are detected. Fragmentation status of the nucleic acid of interest is determined, coincident detection of signals from two or more of the different probes indicating the nucleic acid of interest is not fragmented between the probes.
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Citations
25 Claims
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1. A method of determining fragmentation status of a nucleic acid of interest in a sample at the start of the method, the method comprising:
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contacting the nucleic acid of interest in a reaction mixture with two or more different probes complementary to sites separated by a point of potential fragmentation, wherein the probes each comprise a detectable marker; flowing the nucleic acid of interest into a detection region; detecting detectable marker signals from the probes; and determining fragmentation status of the nucleic acid of interest, wherein coincident detection of signals from two or more of the different probes indicates the nucleic acid of interest was not fragmented between the probes at the start of the method and has at least a given length, and wherein detecting a signal from only one of the different probes indicates the nucleic acid of interest was fragmented. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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Specification