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HELPER OLIGONUCLEOTIDE FOR IMPROVED EFFICIENCY OF AMPLIFICATION AND DETECTION/QUANTITATION OF NUCLEIC ACIDS

  • US 20180037945A1
  • Filed: 07/26/2017
  • Published: 02/08/2018
  • Est. Priority Date: 08/02/2016
  • Status: Active Grant
First Claim
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1. A method for detecting and/or quantitating a target nucleic acid in a sample comprising:

  • (a) contacting nucleic acids in the sample with amplification reagents, the amplification reagents comprising;

    at least an enzyme comprising DNA polymerase activity;

    at least nucleoside triphosphate monomers or other nucleoside monomers;

    at least one forward primer specific for the target nucleic acid, or at least one reverse primer specific for the target nucleic acid, or a combination thereof, for generating at least one amplicon;

    at least one non-extending helper oligonucleotide, wherein;

    (i) the non-extending helper oligonucleotide does not extend, (ii) the non-extending helper oligonucleotide anneals to part of the same portion of the target nucleic acid as at least one of the primers, and (iii) the non-extending helper oligonucleotide enhances the activity of at least one of the primers; and

    at least one detectable probe specific for the amplicon or at least one DNA binding dye;

    (b) incubating the nucleic acids with the amplification reagents for a period of time and under conditions sufficient for an amplification reaction to occur; and

    (c) detecting the amplicon with the at least one detectable probe or the at least one DNA binding dye.

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