Modulation of DNA polymerase function for DNA diagnostics

We reported on two approaches to modulate and increase single nucleotide discrimination of DNA polymerase for applications in DNA diagnostics using PCR. We show that certain chemical modifications in primer probes are able to increase the discrimination in favor of extension of canonical primer probes versus mismatched primer probes. In a second approach to modulate DNA polymerases in this respect we employed directed combinatorial enzyme design to identify enzymes with increased mismatch extension fidelity.