Hot Start is a well established and convenient method to improve specificity of PCR.
- Hot Start/Cold Stop™ technology requires no enzyme activation step
- Ideal for fast PCR protocols
- Convenient room temperature reaction set-up
- Highly specific amplification
- Minimal optimization with self adjusting Mg₂+ buffer technology
Conventional technologies like antibody mediated inhibition or chemical blocking of DNA polymerases carry a few limitations: long initial activation steps may be required, the performance of the DNA polymerase can be reduced or specificity control may be compromised.
The HotMaster Taq DNA Polymerase uses an innovative technology to achieve Hot Start PCR. A thermostable inhibitor (patent-pending) of the Taq DNA Polymerase is released at high temperatures, thereby immediately activating the enzyme. The inhibitor binds and blocks the substrate-binding site of HotMaster Taq in a temperature-dependent manner. At tempertures above 60°C, the inhibitor is released and the HotMaster Taq becomes activated (Hot Start).
At temperatures below 60°C, the inhibitor inactivates the Taq polymerase (Cold Stop), preventing non-specific primer annealing or primer-dimer formation. Unlike many conventional Hot Start PCR techniques, such binding-releasing equilibrium is reversible and can go through all the PCR cycle. This innovative Hot Start/Cold Stop technology provides the ultimate temperature control from the FIRST to the LAST PCR cycles and eliminates the unwanted non-specific amplicons.
Ordering information: HotMaster Taq DNA Polymerase is packaged with 10X HotMaster Taq Buffer with 25 mM Mg2+.
HotMasterMix is a 2.5x ready-to-use format with final concentrations of 1.0 U/50 µl Taq DNA Polymerase, 45 mM KCl, 2.5 mM Mg2+, and 200 µM of each dNTP, in a 50 µl PCR reaction.
