Why use a hot-start Taq?
As PCR reactions sit at room temperature, during assay setup, nonspecific amplification can occur via:
In hot-start PCR, Taq polymerase is inactive until heated. Hot-start PCR activation approaches allow users to minimize non-specific amplification while increasing target yield and specificity.
How does our hot start technology work?
Our JumpStart Taq DNA Polymerase is an antibody inactivated hot-start enzyme. During the initial denature PCR step, Taq DNA Polymerase activity is restored. The resulting PCR exhibits a higher specificity and yield. Unlike chemically modified hot-starts that can take up to 10 min for enzyme activation, antibody mediated hot-start enzymes are activated within 1 min.
Mechanism of antibody mediated hot start PCR
Note: that Magnesium Chloride is added separately if not already in the PCR buffer or when previous optimisation has revealed a requirement for a concentration.
b. Combine reaction components into a 1.5 mL microcentrifuge tube on ice