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Processes for detecting or quantifying nucleic acids using an array of fixed or immobilized nucleic acids

  • US 9,650,666 B2
  • Filed: 07/29/2015
  • Issued: 05/16/2017
  • Est. Priority Date: 06/30/2001
  • Status: Expired due to Term
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 and a second set of primers, wherein the second set of primers comprises at least one production center;

    (iv) polymerizing means for synthesizing nucleic acid copies of said nucleic acid analytes using the first set of primers and the second set of primers;

    (v) a set of oligonucleotides or polynucleotides complementary to at least one segment or sequence of said second set of primers;

    (vi) ligating means for ligating said set of oligonucleotides or polynucleotides (v); and

    (vii) synthesizing means for synthesizing nucleic acid copies under isothermal or isostatic conditions from a production center;

    b) contacting said library of nucleic acid analytes with said first set of primers to form more than one first bound entity, each comprising a first set primer hybridized to a nucleic acid analyte;

    c) using said polymerizing means, extending said bound first set primers of the first bound entities using said nucleic acid analytes of the first bound entities as templates to form first copies of said analytes;

    d) using said ligating means, ligating said set of oligonucleotides or polynucleotides a)(v) to the 3′

    end of said first copies formed in step c) to form more than one ligated product;

    e) contacting said ligated products with said second set of primers to form more than one second hound entity, each comprising a second set primer hybridized to one of the ligated products;

    f) using said polymerizing means, extending said bound second set of primers of the second bound entities using said ligated products of the second bound entities as templates to form more than one complex comprising said ligated products and said extended second set primers;

    g) using said synthesizing means, synthesizing from a production center in said second set primers incorporated in said complexes one or more nucleic acid copies under isothermal or isostatic conditions;

    h) hybridizing said nucleic acid copies formed in step g) to said array of nucleic acids provided in step a)(i); and

    i) detecting or quantifying any of said hybridized copies obtained in step h).

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