Patent ID: 6667165
Filing Date: 2003-12-23
Classification: C12N,C12Q

Abstract:
A method of polynucleotide synthesis comprising:combining in a polymerization reaction mixture a thermostable DNA polymerase from a thermophilic Eubacteria or Archaebacteria or a mixture thereof, a template nucleic acid molecule, appropriate primers for the template nucleic acid molecule, at least one deoxynucleoside triphosphate, and a non-nucleic acid polyanion, wherein the temperature of the polymerization reaction mixture is both sufficient to support some level of thermostable DNA polymerase activity and is such that the non-nucleic acid polyanion inhibits thermostable DNA polymerase activity; heating the polymerization reaction mixture to a temperature at which the template nucleic acid molecule is denatured from a double-stranded molecule to a single-stranded molecule; cooling the polymerization reaction mixture to a temperature at which appropriate primers anneal to the single-stranded molecule; and modifying the temperature of the polymerization reaction mixture to a temperature at which the non-nucleic acid polyanion is substantially dissociated from the thermostable DNA polymerase, wherein the thermostable DNA polymerase recognizes and provides polynucleotide synthesis on primer annealed nucleic acid molecule.