Patent ID: 8592148

Claim:
A method for detecting a catalytic event having improved binding kinetics, said method comprising: (a) immobilizing a target nucleic acid, a primer nucleic acid, and a polymerase onto a surface; (b) contacting said surface with a plurality of nanoparticles, wherein each nanoparticle comprises a plurality of nucleotide phosphates (NPs), which are substrates for the polymerase, wherein each of said plurality of nucleotide phosphates (NPs) is attached to the nanoparticle via a linker, wherein the nanoparticle size, the linker and the NP density on the nanoparticle surface are selected such that the K m for the binding of nanoparticle-immobilized NPs to the polymerase is lowered compared to the K m for a free nucleotide in solution binding to the polymerase; and (c) detecting polymerase-mediated incorporation of at least one of said plurality of nucleotide phosphates into a single molecule of the primer nucleic acid to detect said catalytic event, wherein the nanoparticle is released from the nucleotide phosphate upon incorporation, and wherein the catalytic event has improved binding kinetics.