Thermostable polymerase inhibitor compositions and methods

A dumbbell aptamer comprising

• a single oligonucleotide in a folded configuration, the single oligonucleotide comprising
• nucleotides of SEQ ID NO:1, or a variant thereof, covalently linked to
• nucleotides of SEQ ID NO:3, or a variant thereof, covalently linked to
• nucleotides of SEQ ID NO:2, or a variant thereof, covalently linked to
• nucleotides of SEQ ID NO:3, or the variant thereof;
• in the folded configuration the single oligonucleotide comprises
• a nucleotide stem and two nucleotide loops,
• wherein the nucleotide stem comprises
• the nucleotides of SEQ ID NO:1, or the variant thereof,
• hybridized to
• the nucleotides of SEQ ID NO:2, or the variant thereof,
• wherein the nucleotides of
• SEQ ID NO:1, or the variant thereof, or
• SEQ ID NO:2, or the variant thereof,
• are split into
• a 5′ portion and
• a 3′ portion
• due to a lack of a covalent bond between adjacent nucleotides of
• SEQ ID NO:1 or the variant thereof, or
• SEQ ID NO:2, or the variant thereof,
• the lack of the covalent bond defining
• a 5′ terminus and
• a 3′ terminus
• of the dumbbell aptamer that are positioned adjacent to one another within the nucleotide stem of the single oligonucleotide in the folded configuration,
• wherein:
• the variant of SEQ ID NO:
• 1 has only 1 to 4 nucleotide substitutions in SEQ ID NO:1;
• the variant of SEQ ID NO:2 has only 1 to 4 nucleotide substitutions in SEQ ID NO:2;
• the variants of SEQ ID NO:
• 1 and SEQ ID NO:2 are
• equal in length to each other, and are
• 100% complementary to each other along their entire length;
• and
• each of the two nucleotide loops of the single oligonucleotide, in the folded configuration, individually comprise
• SEQ ID NO:3, or variants thereof,
• the variants of SEQ ID NO:3 having only 1 or 2 nucleotide substitutions in SEQ ID NO:
• 3.