Detection and quantification process for more than one nucleic acid in library
First Claim
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1. A process for detecting or quantifying more than one nucleic acid of interest in a library comprising the steps of:
- a) providing;
(i) an array of fixed or immobilized nucleic acids complementary to said nucleic acids of interest;
(ii) a library of nucleic acid analytes which may contain the nucleic acids of interest sought to be detected or quantified;
(iii) means for attaching one or more non-inherent heteropolymeric universal detection targets (UDT) to a nucleic acid, wherein said attaching means is selected from blunt-end ligation with DNA ligase, ligation of single-stranded ends with RNA ligase or a combination thereof;
(iv) a universal detection element (UDE) which generates a signal directly or indirectly;
b) attaching said non-inherent UDTs to said library of nucleic acid analytes;
c) hybridizing said library of nucleic acid analytes from step b) with said array of fixed or immobilized nucleic acids to form hybrids if said nucleic acids of interest are present;
d) contacting said UDE with said non-inherent UDTs to form a complex bound to said array; and
e) detecting or quantifying said more than one nucleic acid of interest by detecting or measuring the amount of signal generated from any UDE bound to said array.
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Abstract
This invention provides novel compositions and processes for analyte detection, quantification and amplification. Nucleic acid arrays and libraries of analytes are usefully incorporated into such compositions and processes. Universal detection elements, signaling entities and the like are employed to detect and if necessary or desirable, to quantify analytes. Amplification of target analytes are also provided by the compositions and processes of this invention.
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Citations
57 Claims
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1. A process for detecting or quantifying more than one nucleic acid of interest in a library comprising the steps of:
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a) providing; (i) an array of fixed or immobilized nucleic acids complementary to said nucleic acids of interest; (ii) a library of nucleic acid analytes which may contain the nucleic acids of interest sought to be detected or quantified; (iii) means for attaching one or more non-inherent heteropolymeric universal detection targets (UDT) to a nucleic acid, wherein said attaching means is selected from blunt-end ligation with DNA ligase, ligation of single-stranded ends with RNA ligase or a combination thereof; (iv) a universal detection element (UDE) which generates a signal directly or indirectly; b) attaching said non-inherent UDTs to said library of nucleic acid analytes; c) hybridizing said library of nucleic acid analytes from step b) with said array of fixed or immobilized nucleic acids to form hybrids if said nucleic acids of interest are present; d) contacting said UDE with said non-inherent UDTs to form a complex bound to said array; and e) detecting or quantifying said more than one nucleic acid of interest by detecting or measuring the amount of signal generated from any UDE bound to said array. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A process for detecting or quantifying more than one nucleic acid of interest in a library comprising the steps of:
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a) providing; (i) an array of fixed or immobilized nucleic acids complementary to said nucleic acids of interest; (ii) a library of nucleic acid analytes which may contain the nucleic acids of interest sought to be detected or quantified; (iii) means for attaching one or more non-inherent heteropolymeric universal detection targets (UDT) to a nucleic acid, wherein said attaching means is selected from blunt-end ligation with DNA ligase, ligation of single-stranded ends with RNA ligase or a combination thereof; (iv) a universal detection element (UDE) which generates a signal directly or indirectly; b) attaching said non-inherent UDTs to said library of nucleic acid analytes; c) contacting said UDE with said non-inherent UDTs in said library of nucleic acid analytes from step b) to form one or more complexes; d) hybridizing said library of nucleic acid analytes from step c) with said array of nucleic acids to form hybrids if said nucleic acids of interest are present; and e) detecting or quantifying said more than one nucleic acid of interest by detecting or measuring the amount of signal generated from any UDE bound to said array. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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37. A process for detecting or quantifying more than one nucleic acid of interest in a library comprising the steps of:
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a) providing; (i) an array of fixed or immobilized nucleic acids complementary to said nucleic acids of interest; (ii) a library of nucleic acid analytes which may contain the nucleic acids of interest sought to be detected or quantified; and (iii) a universal detection element (UDE) which binds to a domain formed by nucleic acid hybrids for complex formation and generates a signal directly or indirectly, wherein said UDE is selected from a protein, an intercalator, and a combination thereof; b) hybridizing said library of nucleic acid analytes with said array of nucleic acids to form hybrids if said nucleic acids of interest are present, wherein said formed hybrids generate a domain for complex formation; c) contacting said UDE with said hybrids to form a complex bound to said array; and d) detecting or quantifying said more than one nucleic acid of interest by detecting or measuring the amount of signal generated from any UDE bound to said array. - View Dependent Claims (38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57)
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Specification