Patent ID: 8715927

Claim:
A method of identifying an agent that inhibits the activity of a DNA polymerase, comprising: (i) mixing a template oligodeoxynucleotide, a primer oligodeoxynucleotide and the DNA polymerase, wherein the template oligodeoxynucleotide comprises in the 5′ to 3′ direction: (a) a 5′-OH, wherein the 5′-OH is phosphorylated or non-phosphorylated; (b) a poly dT or a poly dA track of about 8 to about 16 nucleotides; (c) a short segment of nucleotides about 3 to about 7 nucleotides in length, wherein the nucleotides include A, C and G when (b) is a poly dT track, or the nucleotides include T, C and G when (b) is a poly dA track; (d) a primer binding sequence that is complementary to the nucleotide sequence of the primer oligodeoxynucleotide, wherein the primer binding sequence comprises the nucleotide sequence of SEQ ID NO: 15; (e) a linker sequence; (f) a biotin-conjugated nucleotide; and (g) 1 to 3 nucleotides at the 3′ terminus; (ii) adding unlabeled dNTPs and a fluorescently-labeled dTTP or dATP; (iii) adding a candidate agent; and (iv) incubating the template oligodeoxynucleotide, the primer oligodeoxynucleotide, the DNA polymerase, the unlabeled dNTPs, the fluorescently-labeled dTTP or dATP, and the candidate agent for a sufficient period of time to allow DNA polymerization, wherein a reduction in incorporation of the fluorescently labeled dTTP or dATP in the presence of candidate agent, relative to incorporation of the fluorescently labeled dTTP or dATP in the absence of the candidate agent, indicates the candidate agent is an agent the inhibits the activity of a DNA polymerase.