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US 6,355,431 B1Granted
Detection of nucleic acid amplification reactions using bead arrays
AssigneeIllumina, Inc.
InventorsMark S. Chee, Kevin Gunderson
Life of this patent
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Abstract
The invention relates to compositions and methods for detecting and quantifying a target nucleic acid using a variety of both signal amplification and target amplification techniques.
Claims35
A method for detecting a first target nucleic acid sequence comprising:
- a) hybridizing at least a first primer nucleic acid to said first target sequence to form a first hybridization complex;
- b) contacting said first hybridization complex with a first enzyme that causes a modification of said first primer nucleic acid to form a modified first primer nucleic acid;
- c) disassociating said first hybridization complex;
- d) contacting said modified first primer nucleic acid with an array comprising:
- i) a substrate with a surface comprising discrete sites;
- and
- ii) a population of microspheres comprising at least a first subpopulation comprising a first capture probe;
- such that said first capture probe and the modified primer form an assay complex;
- wherein said microspheres are randomly distributed on said surface;
- and
- e) detecting the presence of the modified primer nucleic acid.
8 more elements
A method for detecting a target nucleic acid sequence comprising:
- a) hybridizing a first primer to a first target sequence to form a first hybridization complex;
- b) contacting said first hybridization complex with a first enzyme to extend said first primer to form a first newly synthesized strand and form a nucleic acid hybrid that comprises an RNA polymerase promoter;
- c) contacting said hybrid with an RNA polymerase that recognizes said RNA polymerase promoter and generates at least one newly synthesized RNA strand;
- d) contacting said newly synthesized RNA strand with an array comprising:
- i) a substrate with a surface comprising discrete sites;
- and
- ii) a population of microspheres comprising at least a first subpopulation comprising a first capture probe;
- such that said first capture probe and the modified primer form an assay complex;
- wherein said microspheres are randomly distributed on said surface;
- and
- e) detecting the presence of the newly synthesized RNA strand.
8 more elements
A kit for the detection of a first target nucleic acid sequence comprising:
- a) at least a first nucleic acid primer substantially complementary to at least a first domain of said target sequence;
- b) at least a first enzyme that will modify said first nucleic acid primer;
- and
- c) an array comprising:
- i) a substrate with a surface comprising discrete sites;
- and
- ii) a population of microspheres comprising at least a first and a second subpopulation, wherein each subpopulation comprises a capture probe;
- wherein said microspheres are randomly distributed on said surface.
5 more elements
Dependencies are inferred from the claim text.