Decoy probes
AssigneeGen-Probe Incorporated
InventorsChristopher C. Adams, Steven T. Brentano, Gary P. Schroth
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Abstract
The present invention features inhibitors of target-independent amplification and the use of such inhibitors for enhancing an amplification protocol. The inhibitors are believed to enhance an amplification protocol by inhibiting the ability of one or more nucleic acid polymerases to use nucleic acid in a polymerase reaction in the absence of target nucleic acid.
Claims31
A purified decoy probe comprising:
- a first nucleotide base recognition sequence region, wherein said first region binds to an RNA polymerase;
- and
- a second nucleotide base recognition sequence region joined directly to the 5′ end of said first region or is joined to the 3′ end or 5′ end of said first region by a non-nucleotide linker, wherein said optionally present second region is present if said first region can be used to produce a functional double-stranded promoter sequence using a complementary oligonucleotide,
- further provided that if said first region is nucleic acid which can be used to produce said functional double-stranded promoter sequence using said complementary oligonucleotide, then said decoy probe does not have a nucleic acid sequence greater than about 10 nucleotides in length joined directly to the 3′ end of said first region and said decoy probe does not have a 3′ end that can participate in a polymerase reaction.
A purified decoy probe comprising:
- a first nucleotide base recognition sequence region, wherein said first region has at least 35% sequence similarity to an RNA polymerase promoter sequence;
- and
- a second nucleotide base recognition sequence region joined directly to the 5′ end of said first region or is joined to the 3′ end or 5′ end of said first region by a non-nucleotide linker, wherein said optionally present second region is present if said first region can be used to produce a functional double-stranded promoter sequence using a complementary oligonucleotide,
- further provided that if said first region is nucleic acid which can be used to produce said functional double-stranded promoter sequence using said complementary oligonucleotide, then said decoy probe does not have a nucleic acid sequence greater than about 10 nucleotides in length joined directly to the 3′ end of said first region and said decoy probe does not have a 3′ end that can participate in a polymerase reaction.
A reagent mixture for use in an amplification reaction comprising an RNA polymerase and a reversible inhibitor of said polymerase, said inhibitor being a decoy probe comprising a nucleotide base recognition sequence having at least 35% sequence similarity to a promoter sequence for said RNA polymerase, wherein said reagent mixture does not contain a nucleic acid substantially complementary to said inhibitor.
Dependencies are inferred from the claim text.