Process for detecting or quantifying nucleic acids in a 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 identical or complementary in part or whole to sequences of 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) a first set of primers, a second set of primers and a third set of primers wherein said third set comprises at least one production center; and
(iv) polymerizing means for synthesizing nucleic acid copies of said nucleic acid analytes using said first, second and third sets of primers; and
b) contacting said library of nucleic acid analytes with said first set of primers to form a first set of bound primers;
c) extending said first set of bound primers by means of template sequences provided by said nucleic acid analytes to form first copies of said analytes;
d) contacting said extended first copies with said second set of primers to form a second set of bound primers;
e) extending said second set of bound primers by means of template sequences provided by said extended first copies to form second copies of said nucleic acid analytes;
f) contacting said second copies with said third set of primers to form more than one third bound entity to form a third set of bound primers;
g) extending said third set of bound primers by means of template sequences provided by said extended second set of primers to form a hybrid comprising a second copy, a third copy and at least one production center;
h) synthesizing from a production center in said third set of primers in said complexes one or more nucleic acid copies under isothermal or isostatic conditions;
i) hybridizing said nucleic acid copies synthesized in step h) to said array of nucleic acids provided in step a) (i); and
j) detecting or quantifying any of said hybridized copies obtained in step i).
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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
35 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 identical or complementary in part or whole to sequences of 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) a first set of primers, a second set of primers and a third set of primers wherein said third set comprises at least one production center; and (iv) polymerizing means for synthesizing nucleic acid copies of said nucleic acid analytes using said first, second and third sets of primers; and b) contacting said library of nucleic acid analytes with said first set of primers to form a first set of bound primers; c) extending said first set of bound primers by means of template sequences provided by said nucleic acid analytes to form first copies of said analytes; d) contacting said extended first copies with said second set of primers to form a second set of bound primers; e) extending said second set of bound primers by means of template sequences provided by said extended first copies to form second copies of said nucleic acid analytes; f) contacting said second copies with said third set of primers to form more than one third bound entity to form a third set of bound primers; g) extending said third set of bound primers by means of template sequences provided by said extended second set of primers to form a hybrid comprising a second copy, a third copy and at least one production center; h) synthesizing from a production center in said third set of primers in said complexes one or more nucleic acid copies under isothermal or isostatic conditions; i) hybridizing said nucleic acid copies synthesized in step h) to said array of nucleic acids provided in step a) (i); and j) detecting or quantifying any of said hybridized copies obtained in step i). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)
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Specification