Absolute PCR Quantification
AssigneeThe Research Foundation for The State University of New York
InventorsVera Gorfinkel, Boris Gorbovitski
Life of this patent
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Abstract
The present application provides methods and devices for absolute quantification of polymerase chain reaction target nucleic acids. In particular, the methods and devices of the present application provide for splitting a nucleic acid sample to be analyzed into small, isolated volumes, conducting the method of polymerase chain reaction (PCR) on said volumes, detecting PCR amplification products, analyzing said detected PCR amplification products, performing absolute quantification of the PCR target and presenting said quantification results.
Claims50
A method of counting target nucleic acid molecules, comprising:
- a) providing a sample containing a plurality of target nucleic acid molecules;
- b) applying said sample to a multiple vessel array comprising a plurality of reaction vessels, wherein each of said reaction vessels receives an essentially equal sub-volume of said sample, wherein the combined volumes of said sub-volumes defines a tested volume, and wherein the number of said plurality of reaction vessels is comparable with the number of target nucleic acid molecules in said tested volume;
- c) treating said multiple vessel array to conditions to produce a detectable signal in the presence of said target nucleic acid;
- and
- d) determining how many of said reaction vessels contain reactions that produce said detectable signal.
16-22. (canceled)
27-39. (canceled)
A method of sampling a target nucleic acid, comprising:
- a) providing a sample tray comprising a plurality of isolated reaction zones, wherein each isolated reaction zone contains a nucleic acid binding agent having a specific nucleic acid binding rate, wherein the binding rates of the binding agents in different isolated reaction zones are variable binding rates;
- b) exposing the sample tray to a sample comprising target nucleic acid under conditions wherein each isolated reaction is exposed to said sample for essentially the same amount of time;
- c) removing unbound target nucleic acid from said sample tray;
- d) detecting the target nucleic acid in said isolated reaction zones.
47-49. (canceled)
A method of sampling a target nucleic acid, comprising:
- a) providing a sample tray comprising a plurality of isolated reaction zones, wherein each isolated reaction zone contains a nucleic acid binding agent having a specific nucleic acid binding capacity, wherein the binding capacities of the binding agents in different isolated reaction zones are variable binding capacities;
- b) exposing the sample tray to a sample comprising target nucleic acid under conditions wherein said binding agent binds to said target nucleic acid;
- c) removing unbound target nucleic acid from said sample tray;
- d) detecting the target nucleic acid in said isolated reaction zones.
54-56. (canceled)
A method of sampling a target nucleic acid, comprising:
- a) providing a sample tray comprising a plurality of isolated reaction zones, wherein each isolated reaction zone defines a specific reaction sub-volume, wherein the reaction sub-volumes of said different isolated reaction zones are variable reaction sub-volumes;
- b) exposing the sample tray to a sample comprising one or more target nucleic acid molecules under conditions wherein said sample fills said plurality of isolated reaction zones;
- c) detecting the target nucleic acid in said isolated reaction zones.
61-63. (canceled)
(canceled)
Dependencies are inferred from the claim text.