A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
Wayne M. Barnes, Zhian Zhang, Milko Kermekchiev
Abstract
) DNA polymerase is a gain of function mutation that supports faster PCR: the extension times for PCR amplification can be 2-3 times shorter. Surprising results from negative controls led to the discovery of strand-displacement ability and reverse transcriptase activity of Taq D732N DNA polymerase. We demonstrate that the mutant enzyme can, by itself, catalyze RT-PCR, and RT-LAMP assays. Residue 732 is on the surface of the enzyme, not near the active site.
Topics & Concepts
Thermus aquaticusMultiple displacement amplificationHot start PCRMolecular biologyDNA polymeraseTaq polymerasePolymeraseReverse transcriptaseBiologyDNA polymerase IChemistryDNAPolymerase chain reactionBiochemistryNested polymerase chain reactionDNA extractionGeneMolecular Biology Techniques and ApplicationsAdvanced biosensing and bioanalysis techniquesBiosensors and Analytical Detection