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Modifications for antisense compounds

  • US 10,087,445 B2
  • Filed: 11/23/2016
  • Issued: 10/02/2018
  • Est. Priority Date: 03/26/2010
  • Status: Active Grant
First Claim
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1. A method of detecting a target oligonucleotide in a sample, the method comprising:

  • (a) contacting the sample with a composition comprising a probe oligonucleotide capable of hybridizing with the target oligonucleotide, the probe oligonucleotide having the structure 5′

    -Y1

    X—

    Y2-3′

    , whereinY1 comprises a fluorophore at or near the 5′

    -terminus of the probe oligonucleotide and a sequence of 8-12 DNA or RNA nucleotides comprising a nucleotide N1 having a 3′

    phosphate covalently linked to X;

    Y2 comprises a quencher located at or near the 3′

    -terminus of the probe oligonucleotide and a sequence of DNA or RNA nucleotides comprising a nucleotide N2 having a 5′

    phosphate covalently linked to X; and

    X comprises an internal quencher;

    wherein fluorescence of the fluorophore is reduced when the probe oligonucleotide is not hybridized to the target oligonucleotide; and

    (b) detecting the presence of the target oligonucleotide in the sample when an increase in fluorescence of the composition is detected as compared to the fluorescence of the composition in a control sample devoid of the target oligonucleotide.

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