×

Methods of analysis of nucleic acids

  • US 20030165865A1
  • Filed: 01/25/2002
  • Published: 09/04/2003
  • Est. Priority Date: 01/29/2001
  • Status: Abandoned Application
First Claim
Patent Images

1. A method for determining polynucleotide expression comprising:

  • a) providing at least one target polynucleotide, said polynucleotide having a 3′

    end and a 5′

    end;

    b) providing a first oligonucleotide primer, wherein a portion of said first primer is capable of hybridizing to said target polynucleotide;

    c) obtaining a first strand cDNA by reverse transcription of said target polynucleotide, said first strand cDNA having a 5′

    end and a 3′

    end, wherein said 5′

    end of said first strand cDNA contains a sequence corresponding to said first oligonucleotide primer and said 3′

    end of said first strand cDNA comprises at least one nucleotide that extends beyond the 5′

    end of said target polynucleotide to provide a single-stranded extension;

    d) providing a second oligonucleotide primer, wherein at least a portion of said second oligonucleotide primer is capable of hybridizing to said single-stranded extension;

    e) extending said first strand cDNA using said second oligonucleotide primer as a template to produce an extended first strand cDNA containing said first oligonucleotide primer and a region complementary to said second oligonucleotide primer;

    f) amplifying said extended first strand cDNA in the presence of at least one detectable label to produce amplified cDNA such that said amplified cDNA contains said at least one detectable label;

    g) digesting said amplified cDNA to produce a digested cDNA;

    g) hybridizing said digested cDNA to a capture probe coupled to a solid particle under stringent conditions wherein said capture probe is specific for said target polynucleotide and said particle identifies said capture probe; and

    h) determining if said digested cDNA has hybridized to said capture probe thereby identifying said target polynucleotide.

View all claims
  • 1 Assignment
Timeline View
Assignment View
    ×
    ×