DETECTION AND QUANTIFICATION OF BIOMOLECULES USING MASS SPECTROMETRY
First Claim
1. A detector oligonucleotide comprising a complementary portion that is complementary to a target nucleic acid, and a non-complementary portion that is susceptible to cleavage by a cleavage agent, thereby producing a mass distinguishable product (MDP), wherein the detector oligonucleotide comprises a modification selected from the group consisting (i) a modification that increases the melting temperature of the detector oligonucleotide, (ii) a modification that increases resistance of the detector oligonucleotide to cleavage, (iii) a modification that makes the 5′
- portion of the detector oligonucleotide less likely to hybridize to the target nucleic acid than without the modification, (iv) a modification of incorporating a capture mechanism in the detector oligonucleotide, (iv) a modification of incorporating a releasing mechanism in the detector oligonucleotide.
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Abstract
The present invention is directed in part to a method for detecting a target nucleic acid using detector oligonucleotides detectable by mass spectrometry. This method takes advantage of the 5′ to 3′ nuclease activity of a nucleic acid polymerase to cleave annealed oligonucleotide probes from hybridized duplexes and releases labels for detection by mass spectrometry. This process is easily incorporated into a polymerase chain reaction (PCR) amplification assay. The method also includes embodiments directed to quantitative analysis of target nucleic acids.
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Citations
20 Claims
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1. A detector oligonucleotide comprising a complementary portion that is complementary to a target nucleic acid, and a non-complementary portion that is susceptible to cleavage by a cleavage agent, thereby producing a mass distinguishable product (MDP), wherein the detector oligonucleotide comprises a modification selected from the group consisting (i) a modification that increases the melting temperature of the detector oligonucleotide, (ii) a modification that increases resistance of the detector oligonucleotide to cleavage, (iii) a modification that makes the 5′
- portion of the detector oligonucleotide less likely to hybridize to the target nucleic acid than without the modification, (iv) a modification of incorporating a capture mechanism in the detector oligonucleotide, (iv) a modification of incorporating a releasing mechanism in the detector oligonucleotide.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
Specification