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 capture mechanism in the detector oligonucleotide on the non-complementary portion that allows for the mass-distinguishable product to bind to a solid support upon its cleavage from the detector oligonucleotide by the cleavage agent.
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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.
64 Citations
17 Claims
- 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 capture mechanism in the detector oligonucleotide on the non-complementary portion that allows for the mass-distinguishable product to bind to a solid support upon its cleavage from the detector oligonucleotide by the cleavage agent.
Specification