Nucleic acid ligands that bind to and inhibit DNA polymerases
AssigneeNeXstar Pharmaceuticals, Inc.
InventorsLarry Gold, Sumedha Javasena
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Abstract
This invention discloses high-affinity oligonucleotide ligands to the thermostable Taq polymerase and Tth polymerase. Specifically, this invention discloses DNA ligands having the ability to bind to the Taq and Tth polymerases and the methods for obtaining such ligands. The ligands are capable of inhibiting polymerases at ambient temperatures.
Claims17
A method for inhibiting the activity of a thermostable DNA polymerase, comprising adding a nucleic acid ligand that inhibits said DNA polymerase to a DNA polymerization reaction which is being maintained at a temperature at or below which said ligand inhibits polymerization.
A purified and isolated non-naturally occurring nucleic acid ligand to a thermostable polymerase.
A purified and isolated non-naturally occurring nucleic acid ligand to a thermostable reverse transcriptase.
A method for performing the Polymerase Chain Reaction (PCR) comprising:
- a) mixing a sample containing a nucleic acid sequence that is to be amplified with primers that are complementary to the sequences that flank the sequence to be amplified, Taq polymerase, and a nucleic acid ligand that inhibits the polymerase at a temperature at or below which said ligand inhibits polymerization, yet allows the polymerase to be activated at elevated temperatures;
- wherein said nucleic acid ligand is selected from the group consisting of the sequences set forth in Table 2 (SEQ ID NOS:7-35), Table 3 (SEQ ID NOS:36-66, 76, 77), Table 4 (SEQ ID NOS:67-73) and Table 5 (SEQ ID NO:74);
- and
- b) performing the standard PCR steps of melting the target nucleic acid, annealing the primers to the target nucleic acid, and synthesizing the target nucleic acid by thermal cycling of the mixture.
A method for performing the Polymerase Chain Reaction (PCR) comprising:
- a) mixing a sample containing a nucleic acid sequence that is to be amplified with primers that are complementary to the sequences that flank the sequence to be amplified, Tth polymerase, and a nucleic acid ligand that inhibits the polymerase at a temperature at or below which said ligand inhibits polymerization, yet allows the polymerase to be activated at elevated temperatures;
- wherein said nucleic acid ligand is selected from the group consisting of the sequences set forth in Table 2 (SEQ ID NOS:7-35), Table 3 (SEQ ID NOS:36-66, 76, 77), Table 4 (SEQ ID NOS:67-73) and Table 5 (SEQ ID NO:74);
- and
- b) performing the standard PCR steps of melting the target nucleic acid, annealing the primers to the target nucleic acid, and synthesizing the target nucleic acid by thermal cycling of the mixture.
A PCR kit comprising a thermostable DNA polymerase and a nucleic acid ligand that inhibits said polymerase at a temperature below which said ligand inhibits polymerization, yet allows the polymerase to be activated at elevated temperatures, wherein said thermostable DNA polymerase is Taq polymerase and wherein said nucleic acid ligand is selected from the group consisting of the sequences set forth in Table 2 (SEQ ID NOS:7-35), Table 3 (SEQ ID NOS:36-66, 76, 77), Table 4 (SEQ ID NOS:67-73) and Table 5 (SEQ ID NO:74).
A PCR kit comprising a thermostable DNA polymerase and a nucleic acid ligand that inhibits said polymerase at a temperature below which said ligand inhibits polymerization, yet allows the polymerase to be activated at elevated temperatures, wherein said thermostable DNA polymerase is Tth polymerase and wherein said nucleic acid ligand is selected from the group consisting of the sequences set forth in Table 2 (SEQ ID NOS:7-35), Table 3 (SEQ ID NOS:36-66, 76, 77), Table 4 (SEQ ID NOS:67-73) and Table 5 (SEQ ID NO:74).
Dependencies are inferred from the claim text.