|Title:|| Nucleic acid ligand inhibitors to DNA polymerases|
|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.|
|Inventor(s):|| Gold; Larry (Boulder, CO), Jayasena; Sumedha D. (Boulder, CO) |
|Assignee:|| NeXstar Pharmaceuticals, Inc. (Boulder, CO) |
|Filing Date:||Jun 07, 1995|
|Claims:||1. 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, a thermostable polymerase, and a single-stranded nucleic acid ligand to the polymerase that is capable of inhibiting said polymerase, wherein the inhibition of the polymerase by said nucleic acid is temperature dependent; |
b) heating said mixture to a temperature at which said nucleic, acid ligand no longer inhibits said polymerase; and
c) 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.
2. A PCR kit comprising a thermostable DNA polymerase and a single-stranded nucleic acid ligand to said thermostable DNA polymerase that inhibits the activity of said polymerase, wherein the inhibition of said polymerase by said nucleic ligand is temperature dependent.
3. The method of claim 1 wherein said thermostable polymerase is selected from Thermus aquaticus (Taq polymerase) or Thermus thermophilus (Tth polymerase).